Selective modulators of mutant LRRK2 proteolysis and associated methods of use

ABSTRACT

Bifunctional compounds, which find utility as modulators of non-receptor Leucine-rich repeat kinase 2 (LRRK2), are described herein. In particular, the bifunctional compounds of the present disclosure contain on one end a moiety that binds to the cereblon E3 ubiquitin ligase and on the other end a moiety which binds LRRK2, such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of target protein. The bifunctional compounds of the present disclosure exhibit a broad range of pharmacological activities associated with degradation/inhibition of target protein. Diseases or disorders that result from aberrant regulation of the target protein are treated or prevented with compounds and compositions of the present disclosure.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present disclosure claims benefit of and priority to U.S.Provisional Application No. 62/992,951, filed 21 Mar. 2020, titledSELECTIVE MODULATORS OF MUTANT LRRK2 PROTEOLYSIS AND ASSOCIATED METHODSOF USE, which is incorporated herein in by reference in its entirety forall purposes.

INCORPORATION BY REFERENCE

All cited references are hereby incorporated herein by reference intheir entirety, including U.S. patent application Ser. No. 14/686,640,filed on Apr. 14, 2015, published as U.S. Patent Application PublicationNo. 2015/0291562; and U.S. patent application Ser. No. 14/792,414, filedon Jul. 6, 2015, published as U.S. Patent Application Publication No.2016/0058872; and U.S. patent application Ser. No. 15/953,108, filed onApr. 13, 2018, published as U.S. Patent Application Publication No.2018/0228907; and International Patent Application No.PCT/US2019/032163, filed May 5, 2019, published as International PatentApplication Publication No. WO2019/222173A1.

FIELD OF THE INVENTION

The description provides hetero-bifunctional compounds comprising atarget protein binding moiety and a E3 ubiquitin ligase binding moiety,and associated methods of use. The bifunctional compounds are useful asmodulators of targeted ubiquitination of leucine-rich repeat kinase 2(LRRK2), which is then degraded and/or inhibited.

BACKGROUND

Most small molecule drugs bind enzymes or receptors in tight andwell-defined pockets. On the other hand, protein-protein interactionsare notoriously difficult to target using small molecules due to theirlarge contact surfaces and the shallow grooves or flat interfacesinvolved. E3 ubiquitin ligases (of which hundreds are known in humans)confer substrate specificity for ubiquitination, and therefore, are moreattractive therapeutic targets than general proteasome inhibitors due totheir specificity for certain protein substrates. The development ofligands of E3 ligases has proven challenging, in part because they mustdisrupt protein-protein interactions. However, recent developments haveprovided specific ligands which bind to these ligases. For example,since the discovery of nutlins, the first small molecule E3 ligaseinhibitors, additional compounds have been reported that target E3ligases.

Cereblon is a protein that in humans is encoded by the CRBN gene. CRBNorthologs are highly conserved from plants to humans, which underscoresits physiological importance. Cereblon forms an E3 ubiquitin ligasecomplex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A),and regulator of cullins 1 (ROC1). This complex ubiquitinates a numberof other proteins. Through a mechanism which has not been completelyelucidated, cereblon ubiquitination of target proteins results inincreased levels of fibroblast growth factor 8 (FGF8) and fibroblastgrowth factor 10 (FGF10). FGF8 in turn regulates a number ofdevelopmental processes, such as limb and auditory vesicle formation.The net result is that this ubiquitin ligase complex is important forlimb outgrowth in embryos. In the absence of cereblon, DDB1 forms acomplex with DDB2 that functions as a DNA damage-binding protein.

Bifunctional compounds such as those described in U.S. PatentApplication Publications 2015/0291562 and 2014/0356322 (incorporatedherein by reference), function to recruit endogenous proteins to an E3ubiquitin ligase for ubiquitination and subsequent degradation in theproteasome degradation pathway. In particular, the publications citedabove describe bifunctional or proteolysis-targeting chimeric (PROTAC®)protein degrader compounds, which find utility as modulators of targetedubiquitination of a variety of polypeptides and other proteins, whichare then degraded and/or inhibited by the bifunctional compounds.

Feucine-rich repeat kinase 2 (FRRK2) is a member of the leucine-richrepeat kinase family and is a large multi-domain protein with anN-terminal armadillo domain, ankryin repeat region, a leucine-richrepeat (ERR) domain, a tandem Roco type GTPase domain, a kinase domaincontaining a DFG-like motif, and a C-terminal WD40 domain. The FRRK2protein is 2527 amino acids and a molecular weight of 280 kDa. Catalyticactivities of FRRK2 are associated with the kinase and GTPase domain,and FRRK2 is a heterodimer in its active form (Greggio E, et al: TheParkinson disease-associated leucine-rich repeat kinase 2 (FRRK2) is adimer that undergoes intramolecular autophosphorylation. J Biol Chem2008, 283:16906-16914). GTP binding is essential for kinase activity,and mutations that prevent GTP binding have been shown to ablate LRRK2kinase activity (Ito G, et al: GTP binding is essential to the proteinkinase activity of LRRK2, a causative gene product for familialParkinson's disease. Biochemistry 2007, 46:1380-1388). The onlyvalidated physiological substrates (other than LRRK2 itself) are asubset of low-molecular weight G-proteins including Rab8a and Rab10,which are involved in regulation of vesicle trafficking and endosomefunction and trafficking on cytoskeletal networks (Steger M, et al:Phosphoproteomics reveals that Parkinson's disease kinase LRRK2regulates a subset of Rab GTPases. Elife 2016, 5. e12813). Expressionlevels of LRRK2 are highest in immune cells (neutrophils, monocytes andB cells), lung and kidney, with lower levels in the brain where it isexpressed in dopaminergic neurons of the substantia nigra (West A B, etal: Differential LRRK2 expression in the cortex, striatum, andsubstantia nigra in transgenic and nontransgenic rodents. J Comp Neurol2014, 522:2465-2480).

There are several dominant gain-of-function pathogenic and characterizedmutations to LRRK2, located either in the Roco domains (N1437H,R1441G/C/H, Y1699C), effecting GTP hydrolysis, or in the kinase domain(G2019S and I2020T). The G2019S is the most common LRRK2 mutation linkedto Parkinson's disease (PD), which is a progressive neurodegenerativedisorder characterized by resting tremors, rigidity, decreased movement(bradykinesia), and postural instability. The histological hallmarks ofPD include neurodegeneration of the dopaminergic neurons in thesubstantia nigra pars compacta as well as intracellular inclusionscalled Lewy bodies and neurites consisting of the aggregated form of thealpha-synuclein protein. G2019S is associated with 1-2% of all PDpatients and causes an increase in kinase activity of 2-fold in vitro(West A B, et al: Parkinson's diseaseassociated mutations inleucine-rich repeat kinase 2 augment kinase activity. Proc Natl Acad SciUSA 2005, 102: 16842-16847) and autophosphorylation at Ser1292 isincreased 4-fold (Sheng Z, et air Ser1292 autophosphorylation is anindicator of LRRK2 kinase activity and contributes to the cellulareffects of PD mutations. Sci Transl Med 2012, 4:164ra161). The G2019Sand I2020T mutations lie within the DFG motif (DYGI in the case ofLRRK2), common to all kinases, which controls catalytic activity. Thesemutations are thought to disrupt the inactive conformation and thusincrease catalytic activity (Schmidt S H, et al: The dynamic switchmechanism that leads to activation of LRRK2 is embedded in the DFGpsimotif in the kinase domain. Proc Natl Acad Sci USA 2019,116:14979-14988). Several of the above Parkinson disease-associatedmutations (R1441C/G, Y1699C and I2020T) suppress phosphorylation ofFRRK2 at Ser910 and Ser935, which in turn reduces LRRK2 association with14-3-3 proteins, thought to represent an inactive form of LRRK2 (NicholsJ, et al: 14-3-3 binding to LRRK2 is disrupted by multiple Parkinson'sdisease associated mutations and regulates cytoplasmic localisation.Biochem J 2010, 430:393-404).

Furthermore, LRRK2 is linked to autosomal dominant inherited PD througha mutation within a region of chromosome 12, termed PARK8, which islinked to the LRRK2 gene (Lunayama M, et al: A new locus for Parkinson'sdisease (PARK8) maps to chromosome 12p11.2-q13.1. Ann Neurol 2002,51:296-301; Zimprich A, et al.: Mutations in LRRK2 causeautosomal-dominant parkinsonism with pleomorphic pathology. Neuron 2004,44:601-607; Paisan-Ruiz C, et al.: Cloning of the gene containingmutations that cause PARK8-linked Parkinson's disease. Neuron 2004,44:595-600). LRRK2 was first described as having a link to autosomaldominant inherited Parkinson's disease in 1978, where it was traced to afamily in Japan (Nukada H, et al: [A big family of paralysis agitans(author's transl)]. Rinsho Shinkeigaku 1978, 18:627-634). The mostcommon pathogenic LRRK2 mutation (G2019S) occurs in 4-8% of familial and1-3% of sporadic PD cases. In addition, the G2019S mutation is commonamong PD patients of select ancestry, with 30-40% of North AfricanBerber and 14% of Jewish patients harboring the mutation.

LRRK2 kinase inhibitors have been proposed as having the potential totreat mutation-driven PD, where there is an increase in LRRK2 activity,such as G2019S, and idiopathic PD, where the activity of LRRK2 isincreased (Chen J, et al: Leucine-rich repeat kinase 2 in Parkinson'sdisease: updated from pathogenesis to potential therapeutic target. EurNeurol 2018, 79:256-265; Alessi D P et al: LRRK2 kinase in Parkinson'sdisease. Science 2018, 360:36-37; Di Maio R, et al: LRRK2 activation inidiopathic Parkinson's disease. Sci Transl Med 2018, 10). Severaltherapeutics are progressing into the clinic, including LRRK2 kinaseinhibitors that will directly affect phosphorylation of downstreamtargets, and oligonucleotides (ASO's) directly infused into the CNS toblock translation of LRRK2 protein, thereby reducing LRRK2 proteinlevels.

Lewy bodies are the main histological hallmark of PD. Lewy bodies arecomposed primarily of alpha-synuclein aggregates, and mutations inalpha-synuclein that increase this aggregation also increase the risk ofdeveloping PD (Meade R M, et al: Alpha-synuclein structure andParkinson's disease lessons and emerging principles. Mol Neurodegener2019, 14. 29-29). Depletion of LRRK2 with ASOs (Zhao H T, et al: LRRK2antisense oligonucleotides ameliorate a-synuclein inclusion formation ina Parkinson's disease mouse model. Molecular therapy. Nucleic acids2017, 8:508-519) and deletion of LRRK2 at a genomic level have beenshown to reduce alpha-synuclein mediated pathology in mouse models of PD(Lin X, et al.: Leucine-rich repeat kinase 2 regulates the progressionof neuropathology induced by Parkinson's-disease-related mutantalpha-synuclein. Neuron 2009, 64:807-827). Mutations increasing LRRK2activity, such as G2019S, increase the aggregation of alpha-synuclein inneurons and mouse models of PD. This increase was reversed with LRRK2kinase inhibitors (Volpicelli-Daley L A, et al. G2019S-LRRK2 ExpressionAugments α-Synuclein Sequestration into Inclusions in Neurons. JNeurosci. 2016 Jul. 13; 36(28):7415-27. doi:10.1523/JNEUROSCI.3642-15.2016). There is some evidence to suggest thatthe G2019S mutant form of LRRK2 is resistant to inhibition by kinaseinhibitors in the CNS, potentially reducing their disease modifyingeffect (Kelly K, et al. The G2019S mutation in LRRK2 imparts resiliencyto kinase inhibition. Exp Neurol. 2018 November; 309:1-13). Even thoughmost cases of PD also have Lewy bodies upon post-mortem examination,Lewy bodies are not present in a high number of LRRK2 G2019S mutationassociated PD cases (Kalia E V, et al.: Clinical correlations with Lewybody pathology in LRRK2-related Parkinson disease. JAMA neurol 2015,72:100-105). In addition to Lewy bodies being a common feature of PD,Tau pathology is also a major feature of LRRK2 mutation carriers atpost-mortem (Henderson M X, et al.: Alzheimer's disease tau is aprominent pathology in LRRK2 Parkinson's disease. Acta NeuropatholCommun 2019, 7. 183-183). In one study, Tau pathology was observed in100% of LRRK2 mutation carriers, thereby highlighting LRRK2 as animportant target linking PD with Tau pathology in the context of PD,though the genetic causal link was not as strong between LRRK2 andprimary tauopathies, such as supranuclear palsy (PSP) and corticobasaldegeneration (CBD) (Ross O A, et al. (2006) Lrrk2 R1441 substitution andprogressive supranuclear palsy. Neuropathol Appl Neurobiol 32(1):23-25;Sanchez-Contreras M, et al. (2017) Study of LRRK2 variation intauopathy: progressive supranuclear palsy and corticobasal degeneration.Mov Disord 32(1): 115-123). A common variation at the LRRK2 locus as agenetic determinant of PSP survival was recently reported (Jabbari E, etal., Common variation at the LRRK2 locus is associated with survival inthe primary tauopathy progressive supranuclear palsy. bioRxiv2020.02.04.932335; doi: https://doi.org/10.1101/2020.02.04.932335). Ithas been reported that increased LRRK2 expression in PSP by expressionquantitative trait loci (eQTL) analysis may result in a reactivemicroglia-induced proinflammatory state which drives ongoingaccumulation of misfolded Tau protein and clinical disease progression.Functional variants of LRRK2 have also been linked to Crohn's Diseaseand leprosy type 1 inflammatory reactions (Hui K Y, et al. Functionalvariants in the LRRK2 gene confer shared effects on risk for Crohn'sdisease and Parkinson's disease. Sci Transl Med. 2018 Jan. 10; 10(423).pii: eaai7795. doi: 10.1126/scitranslmed.aai7795; Fava et al.Pleiotropic effects for Parkin and LRRK2 in leprosy type-1 reactions andParkinson's disease. Proc Natl Acad Sci USA. 2019 Jul. 30; 116(31):15616-15624. doi: 10.1073/pnas.1901805116. Epub 2019 Jul. 15).

LRRK2 is highly expressed in the immune system in neutrophils, monocytesand macrophages, as well as in brain microglia, and is a modulator ofthe intrinsic regulation of microglial activation and of lysosomaldegradation processes (Ma et al. Genetic comorbidities in Parkinson'sdisease. Hum Mol Genet. 2014 Feb. 1; 23(3):831-41. doi:10.1093/hmg/ddt465. Epub 2013 Sep. 20, which was reviewed in Schapanskyet al. The complex relationships between microglia, alpha-synuclein, andLRRK2 in Parkinson's disease. Neuroscience. 2015 Aug. 27; 302:74-88.doi: 10.1016/j.neuroscience.2014.09.049. Epub 2014 Oct. 2). Prolongedactivation of these immune cells through PD disease processes ormutations in LRRK2 could increase neuroinflammation and lead to agreater risk of developing PD and/or Tau pathology. Treatment withanti-TNF agents reduces the risk of developing PD by 78% in patientswith inflammatory bowel disorder (Peter I, et al.: Anti-tumor necrosisfactor therapy and incidence of Parkinson disease among patients withinflammatory bowel disease. JAMA Neurol 2018), thereby demonstrating thestrong linkage between inflammation and PD. In addition to PD, LRRK2 hasbeen linked to other diseases such as cancer, leprosy, and Crohn'sdisease (Lewis P A, Manzoni C. LRRK2 and human disease: a complicatedquestion or a question of complexes? (2012). Sci Signal. 5(207), pe2).

As such, a class of small molecules targeting LRRK2 are reported to beuseful in the modulation of the LRRK2 protein, including LRRK2 proteinhaving a G2019S mutation (International Patent Application PublicationWO2019/222173A1, incorporated herein by reference).

An ongoing need exists in the art for effective treatments for LRRK2related disease and discorders, e.g., idiopathic PD, LRRK2mutation-associated PD (e.g., PD associated with one or more LRRK2activating mutations), primary tauopathies (e.g., supranuclear palsy(PSP) or corticobasal degeneration (CBD)), lewy body dementia, Crohn'sDisease, Leprosy (e.g., Leprosy with type 1 inflammatory reactions),and/or neuroinflammation.

SUMMARY

The present disclosure describes hetero-bifunctional compounds thatfunction to selectively or preferentially recruit a mutated Leucine-richrepeat kinase 2 (LRRK2) to an E3 ubiquitin ligase as compared to wildtype LRRK2, for targeted ubiquitination and subsequent proteasomaldegradation, and methods of making and using the same. In particular,compounds as described herein preferentially bind to LRRK2 proteinscomprising a G2019S mutation. In addition, the description providesmethods of using an effective amount of a compound of the invention, asdescribed herein, for the treatment or amelioration of a diseasecondition or one or more symptoms thereof, such as a neurodegenerativedisease, e.g., Parkinson's disease, Parkinson disease with dementia,Parkinson's disease at risk syndrome, dementia with Lewy bodies, Lewybody variant of Alzheimer's disease, combined Parkinson's disease andAlzheimer's disease, multiple system atrophy, striatonigraldegeneration, olivopontocerebellar atrophy, and Shy-Drager syndrome,among others.

As such, in one aspect the disclosure provides hetero-bifunctionalcompounds that comprise an E3 ubiquitin ligase binding moiety (i.e., aligand for an E3 ubiquitin ligase (a “ULM” group)), and a proteintargeting moiety that preferentially binds to a mutated form of LRRK2having a G2019S mutation compared to lesser (or no) binding to wild-typeLRRK2 such that the mutated LRRK2 protein is thereby preferentiallyplaced in proximity to the ubiquitin ligase to effect ubiquitination andsubsequent preferential degradation (and/or inhibition) of the mutatedLRRK2 protein. In a preferred embodiment, the ULM (ubiquitin ligasebinding moiety) is a cereblon E3 ubiquitin ligase binding moiety (CLM).For example, the structure of the bifunctional compound can be depictedas:

The respective positions of the PTM and ULM moieties (e.g., CLM), aswell as their number as illustrated herein, is provided by way ofexample only and is not intended to limit the compounds in any way. Aswould be understood by the skilled artisan, the bifunctional compoundsas described herein can be synthesized such that the number and positionof the respective functional moieties can be varied as desired.

In certain embodiments, the bifunctional compound further comprises achemical linker (“L”). In this example, the structure of thebifunctional compound can be depicted as:

where PTM is a moiety that selectively or preferentially binds to anLRRK2 protein having at least one mutation that is a G2019S mutationcompared to binding of the PTM to the wild-type LRRK2, L is a linker,e.g., a bond or a chemical linking group coupling PTM to ULM, and ULM isa cereblon E3 ubiquitin ligase binding moiety (CLM).

For example, the structure of the bifunctional compound can be depictedas:

wherein: PTM is a moiety that selectively or preferentially binds to anLRRK2 protein having at least one mutation that is a G2019S mutationcompared to binding of the PTM to the wild-type LRRK2; “L” is a linker(e.g. a bond or a chemical linking group) coupling the PTM and CLM; andCLM is cereblon E3 ubiquitin ligase binding moiety that binds tocereblon.

In certain embodiments, the compounds as described herein comprisemultiple independently selected ULMs, multiple PTMs, multiple chemicallinkers or a combination thereof.

In any of the aspects or embodiments described herein, the PTM is asmall molecule that selectively or preferentially binds to a LRRK2protein having at least one mutation that is a G2019S mutation comparedto the PTM binding to a wildtype LRRK2. In any of the aspects orembodiments described herein, the PTM is a small molecule capable ofselectively binding the LRRK2 protein having at least one mutation thatis a G2019S mutation, wherein selectivity towards the LRRK2 proteinhaving at least one mutation that is a G2019S mutation is at least 1-60times (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 times)compared to the wild-type LRRK2. In any of the aspects or embodimentsdescribed herein, the PTM is a small molecule that binds the LRRK2protein having at least one mutation that is a G2019S mutation, whereinselectivity towards the LRRK2 protein having at least one mutation thatis a G2019S mutation is at least 1-1000 times (e.g., 50, 100, 200, 300,400, 500, 600, 700, 800, 900 times) compared to the wild-type LRRK2.

In an embodiment, the CLM comprises a chemical group derived from animide, a thioimide, an amide, or a thioamide. In a particularembodiment, the chemical group is a phthalimido group, or an analog orderivative thereof. In a certain embodiment, the CLM is selected fromthalidomide, lenalidomide, pomalidomide, analogs thereof, isosteresthereof, and derivatives thereof. Other contemplated CLMs are describedin U.S. Patent Application Publication No. 2015/0291562, which isincorporated herein by reference in its entirety.

In certain embodiments, “L” is a bond. In additional embodiments, thelinker “L” is a connector with a linear non-hydrogen atom number in therange of 1 to 20. The connector “L” can contain, but is not limited toone or more functional groups such as ether, amide, alkane, alkene,alkyne, ketone, hydroxyl, carboxylic acid, thioether, sulfoxide, andsulfone. The linker can contain aromatic, heteroaromatic, cyclic,bicyclic or tricyclic moieties. Substitution with halogen, such as Cl,F, Br and I can be included in the linker. In the case of fluorinesubstitution, single or multiple fluorines can be included.

In certain embodiments, CLM is a derivative of piperidine-2,6-dione,where piperidine-2,6-dione can be substituted at the 3-position, and the3-substitution can be bicyclic hetero-aromatics with the linkage as C—Nbond or C—C bond. Examples of CLM can be, but are not limited to,pomalidomide, lenalidomide and thalidomide and their analogs.

In an additional aspect, the description provides therapeuticcompositions comprising an effective amount of a compound as describedherein, or a salt form thereof, and a pharmaceutically acceptablecarrier. The therapeutic compositions can be used to trigger targeteddegradation and/or inhibition of a LRRK2 protein that has at least onemutation that is a G2019S mutation in a patient or subject in needthereof, for example, an animal such as a human, and can be used fortreating or ameliorating one or more disease states, conditions, orsymptoms causally related to the mutated LRRK2, which treatment isaccomplished through preferential degradation of the mutated LRRK2protein to control, stabilize or lower levels of protein of the mutatedLRRK2 protein in a patient or subject. In certain embodiments, thetherapeutic compositions as described herein may be used to effectuatethe preferential degradation of mutated LRRK2 (e.g., G2019S) for thetreatment or amelioration of a disease, disorder or symptom, such as,e.g., an infection, an inflammatory or immunological disorder, orcancer.

In yet another aspect, the present disclosure provides a method ofubiquitinating a mutant LRRK2 in a cell. In certain embodiments, themethod comprises administering a hetero-bifunctional compound asdescribed herein comprising a PTM that selectively or preferentiallybinds to a mutated LRRK2 as described herein, and a CLM, preferablylinked together through a chemical linker moiety, as described herein,to effectuate selective or preferential degradation of the mutant LRRK2protein. Though not wanting to be limited by theory, the inventorsbelieve that, pursuant to the invention, poly-ubiquitination of themutated LRRK2 protein will occur when it is placed in proximity to theE3 ubiquitin ligase via use of the hetero-bifunctional compound, therebytriggering subsequent selective or preferential degradation of themutated LRRK2 protein via the proteasomal pathway, thereby controllingor reducing mutant LRRK2 protein levels in cells of the subject. Thecontrol or reduction in mutant LRRK2 protein levels afforded by thepresent disclosure provides treatment of a disease state, condition orat least one causally related symptom, as modulated through a loweringor stabilization of the amount of mutated LRRK2 protein in cells of thesubject.

In still another aspect, the description provides methods for treatingor ameliorating a disease, condition, or symptom thereof in a subject ora patient, e.g., an animal such as a human, comprising administering toa subject in need thereof a composition comprising an effective amount,e.g., a therapeutically effective amount, of a hetero-bifunctionalcompound as described herein or salt form thereof, and apharmaceutically acceptable carrier, wherein the composition iseffective for treating or ameliorating the disease or disorder orsymptom thereof in the subject.

In another aspect, the description provides methods for identifying theeffects of the selective or preferential degradation of a mutant LRRK2protein according to the disclosure in a biological system usingcompounds according to the present disclosure.

In another aspect, the description provides processes and intermediatesfor making a hetero-bifunctional compound of the invention capable oftargeted ubiquitination and selective or preferential degradation of amutant LRRK2 protein according to the disclosure in a cell.

The preceding general areas of utility are given by way of example onlyand are not intended to be limiting on the scope of the presentdisclosure and appended claims. Additional objects and advantagesassociated with the compositions, methods, and processes of the presentdisclosure will be appreciated by one of ordinary skill in the art inlight of the instant claims, description, and examples. For example, thevarious aspects and embodiments of the disclosure may be utilized innumerous combinations, all of which are expressly contemplated by thepresent description. These additional aspects and embodiments areexpressly included within the scope of the present disclosure. Thepublications and other materials used herein to illuminate thebackground of the disclosure, and in particular cases, to provideadditional details respecting the practice, are incorporated byreference.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated into and form a partof the specification, illustrate several embodiments of the presentdisclosure and, together with the description, serve to explain theprinciples of the disclosure. The drawings are only for the purpose ofillustrating embodiments of the disclosure and are not to be construedas limiting the disclosure. Further objects, features and advantages ofthe disclosure will become apparent from the following detaileddescription taken in conjunction with the accompanying figures showingillustrative embodiments of the disclosure.

FIGS. 1A and 1B. Illustration of general principle for the functioningof hetero-bifunctional protein degrading compounds as described herein.FIG. 1A. Exemplary hetero-bifunctional protein degrading compoundscomprise a protein targeting moiety (PTM; darkly shaded rectangle), aubiquitin ligase binding moiety (ULM; lightly shaded triangle), andoptionally a linker moiety (L; black line) coupling or tethering the PTMto the ULM. FIG. 1B Illustrates the functional use of thehetero-bifunctional protein degrading compounds (commercially known asPROTAC® brand compounds) as described herein. Briefly, the ULM(triangle) recognizes and binds to a specific E3 ubiquitin ligase, andthe PTM (large rectangle) binds and recruits a target protein bringingit into close proximity to the E3 ubiquitin ligase. Typically, the E3ubiquitin ligase is complexed with an E2 ubiquitin-conjugating protein(E2), and either alone or via the E2 protein catalyzes attachment ofmultiple ubiquitin molecules (black circles) to a lysine on the targetprotein via an isopeptide bond. The poly-ubiquitinated protein (farright) has thereby been targeted for degradation by the proteosomalmachinery of the cell.

DETAILED DESCRIPTION

The following is a detailed description provided to aid those skilled inthe art in practicing the present invention. Those of ordinary skill inthe art may make modifications and variations in the embodimentsdescribed herein without departing from the spirit or scope of thepresent disclosure. All publications, patent applications, patents, andother references mentioned herein are expressly incorporated byreference in their entirety.

Presently described are compounds, compositions and methods that relateto the surprising and unexpected discovery that an E3 ubiquitin ligase(e.g., a cereblon E3 ubiquitin ligase) ubiquitinates the human LRRK2protein once the E3 ubiquitin ligase and the LRRK2 protein are placed inproximity via a bifunctional compound that binds both the E3 ubiquitinligase and the LRRK2 protein. Accordingly the present disclosureprovides compounds and compositions comprising an E3 ubiquitin ligasebinding moiety (“ULM”) coupled by a bond or chemical linking group (L)to a protein targeting moiety (“PTM”) that targets the LRRK2 protein,which results in the ubiquitination of the LRRK2 protein, and whichleads to degradation of the LRRK2 protein by the proteasome (see FIG. 1).

In one aspect, the description provides compounds in which the PTMpreferably binds the LRRK2 protein. The present disclosure also providesa library of compositions and the use thereof to produce targeteddegradation of the LRRK2 protein in a cell.

In certain aspects, the present disclosure provides hetero-bifunctionalcompounds which comprise a ligand, e.g., a small molecule ligand (i.e.,having a molecular weight of below 2,000, 1,000, 500, or 200 Daltons),which is capable of binding to an E3 ubiquitin ligase, such as cereblon.The compounds also comprise a small molecule moiety that is capable ofbinding to LRRK2 in such a way that the LRRK2 protein is placed inproximity to the ubiquitin ligase to effect ubiquitination anddegradation (and/or inhibition) of the LRRK2 protein. “Small molecule”means, in addition to the above, that the molecule is non-peptidyl, thatis, it is not considered a peptide, e.g., comprises fewer than 4, 3, or2 amino acids. In accordance with the present description, each of thePTM, ULM and hetero-bifunctional molecule is a small molecule.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. The terminology used in thedescription is for describing particular embodiments only and is notintended to be limiting of the disclosure.

Where a range of values is provided, it is understood that eachintervening value in the range, to the tenth of the unit of the lowerlimit unless the context clearly dictates otherwise (such as in the caseof a group containing a number of carbon atoms in which case each carbonatom number falling within the range is provided), between the upper andlower limit of that range and any other stated or intervening value inthat stated range is encompassed within the disclosure. The upper andlower limits of these smaller ranges may independently be included inthe smaller ranges and are also encompassed within the disclosure,subject to any specifically excluded limit in the stated range. Wherethe stated range includes one or both of the limits, ranges excludingeither/or both of those included limits are also included in thedisclosure.

The following terms are used to describe the present disclosure. Ininstances where a term is not specifically defined herein, that term isgiven an art-recognized meaning by those of ordinary skill applying thatterm in context to its use in describing the present disclosure.

The articles “a” and “an” as used herein and in the appended claims areused herein to refer to one or to more than one (i.e., to at least one)of the grammatical object of the article unless the context clearlyindicates otherwise. By way of example, “an element” means one elementor more than one element, unless otherwise indicated.

In the claims, as well as in the specification above, all transitionalphrases such as “comprising,” “including,” “carrying,” “having,”“containing,” “involving,” “holding,” “composed of,” and the like are tobe understood to be open-ended, i.e., to mean including but not limitedto. Only the transitional phrases “consisting of” and “consistingessentially of” shall be closed or semi-closed transitional phrases,respectively, as set forth in the United States Patent Office Manual ofPatent Examining Procedures, Section 2111.03.

It should also be understood that, in certain methods or processesdescribed herein that include more than one step or act, the order ofthe steps or acts of the method is not necessarily limited to the orderin which the steps or acts of the method are recited unless the contextindicates otherwise.

The terms “co-administration” and “co-administering” or “combinationtherapy” refer to both concurrent administration (administration of twoor more therapeutic agents at the same time) and time-variedadministration (administration of one or more therapeutic agents at atime different from that of the administration of an additionaltherapeutic agent or agents), as long as the two or more therapeuticagents are present in the patient to some extent, preferably ateffective amounts, at the same time. In certain preferred aspects, oneor more of the hetero-bifunctional compounds described herein arecoadministered with at least one additional bioactive agent, e.g., ananticancer agent. In particularly preferred aspects, theco-administration of such compounds results in synergistic activityand/or therapy such as, e.g., anticancer activity.

The term “compound”, as used herein, unless otherwise indicated, refersto any specific hetero-bifunctional compound disclosed herein,pharmaceutically acceptable salts and solvates thereof, and deuteratedforms of any of the aforementioned molecules, where applicable.Deuterated compounds contemplated are those in which one or more of thehydrogen atoms contained in the drug molecule have been replaced bydeuterium. Such deuterated compounds preferably have one or moreimproved pharmacokinetic or pharmacodynamic properties (e.g., longerhalf-life) compared to the equivalent “undeuterated” compound.

The term “ubiquitin ligase” refers to a family of proteins thatfacilitate the transfer of one or more ubiquitins to a specificsubstrate protein. Addition of a chain of several ubiquitins(poly-ubiquitination) targets the substrate protein for degradation. Forexample, cereblon is an E3 ubiquitin ligase that alone, or incombination with an E2 ubiquitin-conjugating enzyme, can ultimatelycause the attachment of a chain of four ubiquitins to a lysine residueon the target protein, thereby targeting the protein for degradation bythe proteasome. The ubiquitin ligase is involved in poly-ubiquitinationsuch that a first ubiquitin is attached to a lysine on the targetprotein; a second ubiquitin is attached to the first; a third isattached to the second, and a fourth is attached to the third. Suchpoly-ubiquitination marks proteins for degradation by the proteasome.

The term “patient” or “subject” is used throughout the specification todescribe an animal, preferably a human or a domesticated animal, to whomtreatment, including prophylactic treatment, with the compositionsaccording to the present disclosure is provided. For treatment of thosediseases, conditions or symptoms that are specific for a specificanimal, such as a human patient, the term “patient” refers to thatspecific animal, including a domesticated animal such as a dog or cat,or a farm animal such as a horse, cow, sheep, etc. In general, in thepresent disclosure, the terms “patient” and “subject” refer to a humanpatient unless otherwise stated or implied from the context of the useof the term.

The terms “effective” and “therapeutically effective” are used todescribe an amount of a compound or composition which, when used withinthe context of its intended use, and either in a single dose or, morepreferably after multiple doses within the context of a treatmentregimen, effects an intended result such as an improvement in a diseaseor condition, or amelioration or reduction in one or more symptomsassociated with a disease or condition. The terms “effective” and“therapeutically effective” subsume all other “effective amount” or“effective concentration” terms, which are otherwise described or usedin the present application.

Compounds and Compositions

In one aspect, the description provides hetero-bifunctional compoundscomprising an E3 ubiquitin ligase binding moiety (“ULM”) that is acereblon E3 ubiquitin ligase binding moiety (a “CLM”), The CLM iscovalently coupled to a protein targeting moiety (PTM) that binds to theprotein, which coupling is either directly by a bond or via a chemicallinking group (L) according to the structure:PTM-L-CLM  (A)wherein L is the bond or chemical linking group, and PTM is a proteintargeting moiety that binds to the protein LRRK2, where the PTM is aLRRK2 targeting moiety. The term CLM is inclusive of all cereblonbinding moieties.

In any of the aspects or embodiments, the CLM demonstrates a halfmaximal inhibitory concentration (IC₅₀) for the E3 ubiquitin ligase(e.g., cereblon E3 ubiquitin ligase) of less than about 200 μM. The IC₅₀can be determined according to any suitable method known in the art,e.g., a fluorescent polarization assay.

In certain embodiments, the hetero-bifunctional compounds describedherein demonstrate an IC₅₀ or a half maximal degradation concentration(DC₅₀) of less than about 100, 50, 10, 1, 0.5, 0.1, 0.05, 0.01, 0.005,0.001 mM, or less than about 100, 50, 10, 1, 0.5, 0.1, 0.05, 0.01,0.005, 0.001 μM, or less than about 100, 50, 10, 1, 0.5, 0.1, 0.05,0.01, 0.005, 0.001 nM, or less than about 100, 50, 10, 1, 0.5, 0.1,0.05, 0.01, 0.005, 0.001 μM.

The term “alkyl” shall mean within its context a linear, branch-chainedor cyclic fully saturated hydrocarbon radical, preferably a C₁-C₁₀,preferably a C₁-C₆, or more preferably a C₁-C₃ alkyl group, which may beoptionally substituted with any suitable functional group or groups.Examples of alkyl groups are methyl, ethyl, n-butyl, sec-butyl, n-hexyl,n-heptyl, n-octyl, n-nonyl, n-decyl, isopropyl, 2-methylpropyl,cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl,cyclopentylethyl, cyclohexylethyl and cyclohexyl, among others. Incertain embodiments, the alkyl group is end-capped with a halogen group(At, Br, Cl, F, or I).

The term “Alkenyl” refers to linear, branch-chained or cyclic C₂-C₁₀(preferably C₂-C₆) hydrocarbon radicals containing at least one C═Cbond.

The term “Alkynyl” refers to linear, branch-chained or cyclic C₂-C₁₀(preferably C₂-C₆) hydrocarbon radicals containing at least one C≡Cbond.

The term “alkylene” when used, refers to a —(CH₂)_(n)— group (n is aninteger generally from 0-6), which may be optionally substituted. Whensubstituted, the alkylene group preferably is substituted on one or moreof the methylene groups with a C₁-C₆ alkyl group (including acyclopropyl group or a t-butyl group), but may also be substituted withone or more halo groups, preferably from 1 to 3 halo groups or one ortwo hydroxyl groups, O—(C₁-C₆ alkyl) groups or amino acid sidechains asotherwise disclosed herein. In certain embodiments, an alkylene groupmay be substituted with a urethane or alkoxy group (or other suitablefunctional group) which may be further substituted with a polyethyleneglycol chain (of from 1 to 10, preferably 1 to 6, or more preferably 1to 4 ethylene glycol units) to which is substituted (preferably, but notexclusively on the distal end of the polyethylene glycol chain) an alkylchain substituted with a single halogen group, preferably a chlorinegroup. In still other embodiments, the alkylene (e.g., methylene) group,may be substituted with an amino acid sidechain group such as asidechain group of a natural or unnatural amino acid, for example,alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine,cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine,isoleucine, lysine, leucine, methionine, proline, serine, threonine,valine, tryptophan or tyrosine.

The term “unsubstituted” shall mean substituted only with hydrogenatoms. A range of carbon atoms which includes C₀ means that carbon isabsent and is replaced with H. Thus, a range of carbon atoms which isC₀-C₆ includes carbons atoms of 1, 2, 3, 4, 5 and 6 and for C₀, H standsin place of carbon.

The term “substituted” or “optionally substituted” shall meanindependently (i.e., where more than one substituent occurs, eachsubstituent is selected independent of another substituent) one or moresubstituents (independently up to five substituents, preferably up tothree substituents, more preferably 1 or 2 substituents on a moiety in acompound according to the present disclosure and may includesubstituents which themselves may be further substituted) at a carbon(or nitrogen) position anywhere on a molecule within context, andincludes as possible substituents hydroxyl, thiol, carboxyl, cyano(C≡N), nitro (NO₂), halogen (preferably, 1, 2 or 3 halogens, especiallyon an alkyl, especially a methyl group such as a trifluoromethyl), analkyl group (preferably, C₁-C₁₀, more preferably, C₁-C₆), aryl(especially phenyl and substituted phenyl, for example benzyl orbenzoyl), alkoxy group (preferably, C₁-C₆ alkyl or aryl, includingphenyl and substituted phenyl), thioether (preferably, C₁-C₆ alkyl oraryl), acyl (preferably, C₁-C₆ acyl), ester or thioester (preferably,C₁-C₆ alkyl or aryl) including alkylene ester (such that attachment ison the alkylene group, rather than at the ester function which ispreferably substituted with a C₁-C₆ alkyl or aryl group), halogen(preferably, F or Cl), amine (including a five- or six-membered cyclicalkylene amine, further including a C₁-C₆ alkyl amine or a C₁-C₆ dialkylamine which alkyl groups may be substituted with one or two hydroxylgroups) or an optionally substituted —N(C₀-C₆ alkyl)C(O)(O—C₁-C₆ alkyl)group (which may be optionally substituted with a polyethylene glycolchain to which is further bound an alkyl group containing a singlehalogen, preferably chlorine substituent), hydrazine, amido, which arepreferably independently substituted with one or two C₁-C₆ alkyl groups(including a carboxamide which is optionally substituted with one or twoC₁-C₆ alkyl groups), alkanol (preferably, C₁-C₆ alkyl or aryl), oralkanoic acid (preferably, C₁-C₆ alkyl or aryl). Substituents accordingto the present disclosure may include, for example —SiR₁R₂R₃ groupswhere each of R₁ and R₂ is as otherwise described herein and R₃ is H ora C₁-C₆ alkyl group, preferably R₁, R₂, R₃ together is a C₁-C₃ alkylgroup (including an isopropyl or t-butyl group). Each of theabove-described groups may be linked directly to the substituted moietyor alternatively, the substituent may be linked to the substitutedmoiety (preferably in the case of an aryl or heteroaryl moiety) throughan optionally substituted —(CH₂)_(m)— or alternatively an optionallysubstituted —(OCH₂)_(m)—, —(OCH₂CH₂)_(m)— or —(CH₂CH₂O)_(m)— group,which may be substituted with any one or more of the above-describedsubstituents. Alkylene groups —(CH₂)_(m)— or —(CH₂)_(n)— groups or otherchains such as ethylene glycol chains, as identified above, may besubstituted anywhere on the chain. Preferred substituents on alkylenegroups include halogen or C₁-C₆ (preferably C₁-C₃) alkyl groups, whichmay be optionally substituted with one or two hydroxyl groups, one ortwo ether groups (O—C₁-C₆ groups), up to three halo groups (preferablyF), or a side chain of an amino acid as otherwise described herein andoptionally substituted amide (preferably carboxamide substituted asdescribed above) or urethane groups (often with one or two C₀-C₆ alkylsubstituents, which group(s) may be further substituted). In certainembodiments, the alkylene group (often a single methylene group) issubstituted with one or two optionally substituted C₁-C₆ alkyl groups,preferably C₁-C₄ alkyl group, most often methyl or O-methyl groups or asidechain of an amino acid as otherwise described herein. In the presentdisclosure, a moiety in a molecule may be optionally substituted with upto five substituents, preferably up to three substituents. Most often,in the present disclosure moieties which are substituted are substitutedwith one or two substituents.

The term “substituted” (each substituent being independent of any othersubstituent) shall also mean within its context of use C₁-C₆ alkyl,C₁-C₆ alkoxy, halogen, amido, carboxamido, sulfone, includingsulfonamide, keto, carboxy, C₁-C₆ ester (oxyester or carbonylester),C₁-C₆ keto, urethane —O—C(O)—NR₁R₂ or —N(R₁)—C(O)—O—R₁, nitro, cyano andamine (especially including a C₁-C₆ alkylene-NR₁R₂, a mono- or di-C₁-C₆alkyl substituted amines which may be optionally substituted with one ortwo hydroxyl groups). Each of these groups contain unless otherwiseindicated, within context, between 1 and 6 carbon atoms. In certainembodiments, preferred substituents will include for example, —NH—,—NHC(O)—, —O—, ═O, —(CH₂)_(m)— (here, m and n are in context, 1, 2, 3,4, 5 or 6), —S—, —S(O)—, SO₂— or —NH—C(O)—NH—, —(CH₂)_(n)OH,—(CH₂)_(n)SH, —(CH₂)_(n)COOH, C₁-C₆ alkyl, —(CH₂)_(n)O—(C₁-C₆ alkyl),—(CH₂)_(n)C(O)—(C₁-C₆ alkyl), —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl),—(CH₂)_(n)C(O)O—(C₁-C₆ alkyl), —(CH₂)_(n)NHC(O)—R₁,—(CH₂)_(n)C(O)—NR₁R₂, —(OCH₂)_(n)OH, —(CH₂O)_(n)COOH, C₁-C₆ alkyl,—(OCH₂)_(n)O—(C₁-C₆ alkyl), —(CH₂O)_(n)C(O)—(C₁-C₆ alkyl),—(OCH₂)_(n)NHC(O)—R₁, —(CH₂O)_(n)C(O)—NR₁R₂, —S(O)₂—R_(S), —S(O)—R_(S)(R_(S) is C₁-C₆ alkyl or a —(CH₂)_(m)—NR₁R₂ group), NO₂, CN or halogen(F, Cl, Br, I, preferably F or Cl), depending on the context of the useof the substituent. R₁ and R₂ are each, within context, H or a C₁-C₆alkyl group (which may be optionally substituted with one or twohydroxyl groups or up to three halogen groups, preferably fluorine). Theterm “substituted” shall also mean, within the chemical context of thecompound defined and substituent used, an optionally substituted aryl orheteroaryl group or an optionally substituted heterocyclic group asotherwise described herein. Alkylene groups may also be substituted asotherwise disclosed herein, preferably with optionally substituted C₁-C₆alkyl groups (methyl, ethyl or hydroxymethyl or hydroxyethyl ispreferred, thus providing a chiral center), a sidechain of an amino acidgroup as otherwise described herein, an amido group as describedhereinabove, or a urethane group O—C(O)—NR₁R₂ group where R₁ and R₂ areas otherwise described herein, although numerous other groups may alsobe used as substituents. Various optionally substituted moieties may besubstituted with 3 or more substituents, preferably no more than 3substituents and preferably with 1 or 2 substituents. It is noted thatin instances where, in a compound at a particular position of themolecule substitution is required (principally, because of valency), butno substitution is indicated, then that substituent is construed orunderstood to be H, unless the context of the substitution suggestsotherwise.

The term “aryl” or “aromatic”, in context, refers to a substituted (asotherwise described herein) or unsubstituted monovalent aromatic radical(e.g., a 5-16 membered ring) having a single ring (e.g., benzene,phenyl, benzyl, or 5, 6, 7 or 8 membered ring) or condensed rings (e.g.,naphthyl, anthracenyl, phenanthrenyl, 10-16 membered ring, etc.) and canbe bound to the compound according to the present disclosure at anyavailable stable position on the ring(s) or as otherwise indicated inthe chemical structure presented. Other examples of aryl groups, incontext, may include heterocyclic aromatic ring systems, “heteroaryl”groups having one or more nitrogen, oxygen, or sulfur atoms in the ring(moncyclic) such as imidazole, furyl, pyrrole, furanyl, thiene,thiazole, pyridine, pyrimidine, pyrazine, triazole, oxazole or fusedring systems such as indole, quinoline, indolizine, azaindolizine,benzofurazan, etc., among others, which may be optionally substituted asdescribed above. Among the heteroaryl groups which may be mentionedinclude nitrogen-containing heteroaryl groups such as pyrrole, pyridine,pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole,triazole, triazine, tetrazole, indole, isoindole, indolizine,azaindolizine, purine, indazole, quinoline, dihydroquinoline,tetrahydroquinoline, isoquinoline, dihydroisoquinoline,tetrahydroisoquinoline, quinolizine, phthalazine, naphthyridine,quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine,imidazotriazine, pyrazinopyridazine, acridine, phenanthridine,carbazole, carbazoline, pyrimidine, phenanthroline, phenacene,oxadiazole, benzimidazole, pyrrolopyridine, pyrrolopyrimidine andpyridopyrimidine; sulfur-containing aromatic heterocycles such asthiophene and benzothiophene; oxygen-containing aromatic heterocyclessuch as furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; andaromatic heterocycles comprising 2 or more hetero atoms selected fromamong nitrogen, sulfur and oxygen, such as thiazole, thiadizole,isothiazole, benzoxazole, benzothiazole, benzothiadiazole,phenothiazine, isoxazole, furazan, phenoxazine, pyrazoloxazole,imidazothiazole, thienofuran, furopyrrole, pyridoxazine, furopyridine,furopyrimidine, thienopyrimidine and oxazole, among others, all of whichmay be optionally substituted.

The term “substituted aryl” refers to an aromatic carbocyclic groupcomprised of at least one aromatic ring or of multiple condensed ringsat least one of which being aromatic, wherein the ring(s) aresubstituted with one or more substituents. For example, an aryl groupcan comprise a substituent(s) selected from: —(CH₂)_(n)OH,—(CH₂)_(n)—O—(C₁-C₆)alkyl, —(CH₂)_(n)—O—(CH₂)_(n)—(C₁-C₆)alkyl,—(CH₂)_(n)—C(O)(C₀-C₆) alkyl, —(CH₂)_(n)—C(O)O(C₀-C₆)alkyl,—(CH₂)_(n)—OC(O)(C₀-C₆)alkyl, amine, mono- or di-(C₁-C₆ alkyl) aminewherein the alkyl group on the amine is optionally substituted with 1 or2 hydroxyl groups or up to three halo (preferably F, Cl) groups, OH,COOH, C₁-C₆ alkyl, preferably CH₃, CF₃, OMe, OCF₃, NO₂, or CN group(each of which may be substituted in ortho-, meta- and/or para-positionsof the phenyl ring, preferably para-), an optionally substituted phenylgroup (the phenyl group itself is preferably connected to a PTM group,including a ULM group, via a linker group), and/or at least one of F,Cl, OH, COOH, CH₃, CF₃, OMe, OCF₃, NO₂, or CN group (in ortho-, meta-and/or para-positions of the phenyl ring, preferably para-), a naphthylgroup, which may be optionally substituted, an optionally substitutedheteroaryl, preferably an optionally substituted isoxazole including amethyl substituted isoxazole, an optionally substituted oxazoleincluding a methyl substituted oxazole, an optionally substitutedthiazole including a methyl substituted thiazole, an optionallysubstituted isothiazole including a methyl substituted isothiazole, anoptionally substituted pyrrole including a methyl substituted pyrrole,an optionally substituted imidazole including a methylimidazole, anoptionally substituted benzimidazole or methoxybenzylimidazole, anoptionally substituted oximidazole or methyloximidazole, an optionallysubstituted diazole group, including a methyldiazole group, anoptionally substituted triazole group, including a methyl substitutedtriazole group, an optionally substituted pyridine group, including ahalo- (preferably, F) or methyl substituted pyridine group or anoxapyridine group (where the pyridine group is linked to the phenylgroup by an oxygen), an optionally substituted furan, an optionallysubstituted benzofuran, an optionally substituted dihydrobenzofuran, anoptionally substituted indole, indolizine or azaindolizine (2, 3, or4-azaindolizine), an optionally substituted quinoline, and combinationsthereof.

“Carboxyl” denotes the group —C(O)OR, where R is hydrogen, alkyl,substituted alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl, whereas these generic substituents have meanings which areidentical with definitions of the corresponding groups defined herein.

The term “heteroaryl” or “hetaryl” can mean but is in no way limited toa 5-16 membered heteroaryl (e.g., 5, 6, 7 or 8 membered monocyclic ringor a 10-16 membered heteroaryl having multiple condensed rings), anoptionally substituted quinoline (which may be attached to thepharmacophore or substituted on any carbon atom within the quinolinering), an optionally substituted indole (including dihydroindole), anoptionally substituted indolizine, an optionally substitutedazaindolizine (2, 3 or 4-azaindolizine) an optionally substitutedbenzimidazole, benzodiazole, benzoxofuran, an optionally substitutedimidazole, an optionally substituted isoxazole, an optionallysubstituted oxazole (preferably methyl substituted), an optionallysubstituted diazole, an optionally substituted triazole, a tetrazole, anoptionally substituted benzofuran, an optionally substituted thiophene,an optionally substituted thiazole (preferably methyl and/or thiolsubstituted), an optionally substituted isothiazole, an optionallysubstituted triazole (preferably a 1,2,3-triazole substituted with amethyl group, a triisopropylsilyl group, an optionally substituted—(CH₂)_(m)—O—C₁-C₆ alkyl group or an optionally substituted—(CH₂)_(m)—C(O)—O—C₁-C₆ alkyl group), an optionally substituted pyridine(2-, 3, or 4-pyridine) or a group according to the chemical structure:

wherein:

-   -   S^(c) is CHR^(SS), NR^(URE), or O;    -   R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally        substituted C₁-C₆ alkyl (preferably substituted with one or two        hydroxyl groups or up to three halo groups (e.g. CF₃),        optionally substituted O(C₁-C₆ alkyl) (preferably substituted        with one or two hydroxyl groups or up to three halo groups) or        an optionally substituted acetylenic group —C≡C—R_(a) where        R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃ alkyl);    -   R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally        substituted C₁-C₆ alkyl (preferably substituted with one or two        hydroxyl groups or up to three halo groups), optionally        substituted O—(C₁-C₆ alkyl) (preferably substituted with one or        two hydroxyl groups or up to three halo groups) or an optionally        substituted —C(O)(C₁-C₆ alkyl) (preferably substituted with one        or two hydroxyl groups or up to three halo groups);    -   R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a        —C(O)(C₁-C₆ alkyl), each of which groups is optionally        substituted with one or two hydroxyl groups or up to three        halogen, preferably fluorine groups, or an optionally        substituted heterocycle, for example piperidine, morpholine,        pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine,        piperazine, each of which is optionally substituted, and    -   Y^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo        (preferably Cl or F), optionally substituted C₁-C₆ alkyl        (preferably substituted with one or two hydroxyl groups or up to        three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆        alkyl) (preferably substituted with one or two hydroxyl groups        or up to three halo groups) or an optionally substituted        acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl        group (preferably C₁-C₃ alkyl).

The terms “aralkyl” and “heteroarylalkyl” refer to groups that compriseboth aryl or, respectively, heteroaryl as well as alkyl and/orheteroalkyl and/or carbocyclic and/or heterocycloalkyl ring systemsaccording to the above definitions.

The term “arylalkyl” as used herein refers to an aryl group as definedabove appended to an alkyl group defined above. The arylalkyl group isattached to the parent moiety through an alkyl group wherein the alkylgroup is one to six carbon atoms. The aryl group in the arylalkyl groupmay be substituted as defined above.

The term “Heterocycle” refers to a cyclic group which contains at leastone heteroatom, e.g., N, O or S, and may be aromatic (heteroaryl) ornon-aromatic. Thus, the heteroaryl moieties are subsumed under thedefinition of heterocycle, depending on the context of its use.Exemplary heteroaryl groups are described hereinabove.

Exemplary heterocyclics include: azetidinyl, benzimidazolyl,1,4-benzodioxanyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl,benzothienyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl,dioxanyl, dioxolanyl, ethyleneurea, 1,3-dioxolane, 1,3-dioxane,1,4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolinyl,imidazolidinyl, indolinyl, indolyl, isoquinolinyl, isothiazolidinyl,isothiazolyl, isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl,oxazolidinyl, oxazolyl, pyridone, 2-pyrrolidone, pyridine, piperazinyl,N-methylpiperazinyl, piperidinyl, phthalimide, succinimide, pyrazinyl,pyrazolinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl,quinolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydroquinoline,thiazolidinyl, thiazolyl, thienyl, tetrahydrothiophene, oxane, oxetanyl,oxathiolanyl, thiane among others.

Heterocyclic groups can be optionally substituted with a member selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SOaryl, —SO— heteroaryl,—SO2-alkyl, —SO2-substituted alkyl, —SO2-aryl, oxo (═O), and—SO2-heteroaryl. Such heterocyclic groups can have a single ring ormultiple condensed rings. Examples of nitrogen heterocycles andheteroaryls include, but are not limited to, pyrrole, imidazole,pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine,isoindole, indole, indazole, purine, quinolizine, isoquinoline,quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline,cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine,phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine,phenothiazine, imidazolidine, imidazoline, piperidine, piperazine,indoline, morpholino, piperidinyl, tetrahydrofuranyl, and the like aswell as N-alkoxy-nitrogen containing heterocycles. The term“heterocyclic” also includes bicyclic groups in which any of theheterocyclic rings is fused to a benzene ring or a cyclohexane ring oranother heterocyclic ring (for example, indolyl, quinolyl, isoquinolyl,tetrahydroquinolyl, and the like).

The term “cycloalkyl” can mean but is in no way limited to univalentgroups derived from monocyclic or polycyclic alkyl groups orcycloalkanes, as defined herein, e.g., saturated monocyclic hydrocarbongroups having from three to twenty carbon atoms in the ring, including,but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl and the like. The term “substituted cycloalkyl” can mean butis in no way limited to a monocyclic or polycyclic alkyl group and beingsubstituted by one or more substituents, for example, amino, halogen,alkyl, substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro,mercapto or sulfo, whereas these generic substituent groups havemeanings which are identical with definitions of the correspondinggroups as defined in this legend.

“Heterocycloalkyl” refers to a monocyclic or polycyclic alkyl group inwhich at least one ring carbon atom of its cyclic structure beingreplaced with a heteroatom selected from the group consisting of N, O, Sor P. “Substituted heterocycloalkyl” refers to a monocyclic orpolycyclic alkyl group in which at least one ring carbon atom of itscyclic structure being replaced with a heteroatom selected from thegroup consisting of N, O, S or P and the group is containing one or moresubstituents selected from the group consisting of halogen, alkyl,substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro, mercapto orsulfo, whereas these generic substituent group have meanings which areidentical with definitions of the corresponding groups as defined inthis legend.

The term “hydrocarbyl” shall mean a compound which contains carbon andhydrogen and which may be fully saturated, partially unsaturated oraromatic and includes aryl groups, alkyl groups, alkenyl groups andalkynyl groups.

The term “independently” is used herein to indicate that the variable,which is independently applied, varies independently from application toapplication.

The term “lower alkyl” refers to methyl, ethyl or propyl

The term “lower alkoxy” refers to methoxy, ethoxy or propoxy.

Exemplary CLMs

Neo-Imide Compounds

In any aspect or embodiment described herein, the description providesCLMs useful for binding and recruiting cereblon. In certain embodiments,the CLM is selected from the group consisting of chemical structures:

wherein:

-   -   W of Formulas (a1) through (e) [e.g., (a1), (a2), (a3), (a4),        (b), (c), (d1), (d2), and/or (e)] is independently selected from        the group CH₂, O, CHR, C═O, SO₂, NH, N, optionally substituted        cyclopropyl group, optionally substituted cyclobutyl group, and        N-alkyl;    -   W₃ of Formulas (a1) through (e) is selected from C or N;    -   X of Formulas (a1) through (e) is independently selected from        the group absent, O, S and CH₂;    -   Y of Formulas (a1) through (e) is independently selected from        the group CH₂, —C═CR′, NH, N-alkyl, N-aryl, N-heteroaryl,        N-cycloalkyl, N-heterocyclyl, O, and S;    -   Z of Formulas (a1) through (e) is independently selected from        the group absent, O, and S or CH₂ except that both X and Z        cannot be CH₂ or absent;    -   G and G′ of Formulas (a1) through (e) are independently selected        from the group H, optionally substituted linear or branched        alkyl, OH, R′OCOOR, R′OCONRR″, CH₂-heterocyclyl optionally        substituted with R′, and benzyl optionally substituted with R′;    -   Q1-Q4 of Formulas (a1) through (e) represent a carbon C or N        substituted with a group independently selected from H, R, N or        N-oxide;    -   A of Formulas (a1) through (e) is independently selected from        the group H, optionally substituted linear or branched alkyl,        cycloalkyl, Cl and F;    -   n of Formulas (a1) through (e) represent an integer from 1 to 10        (e.g., 1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);    -   R of Formulas (a1) through (e) comprises, but is not limited to:        a bond, H, —C(═O)R′ (e.g., a carboxy group), —CONR′R″ (e.g., an        amide group), —OR′ (e.g., OH), —NR′R″ (e.g., an amine group),        —SR′, —SO₂R′, —SO₂NR′R″, —CR′R″—, —CR′NR′R″—, (—CR′O)_(n)R″,        optionally substituted heterocyclyl, optionally substituted        aryl, (e.g., an optionally substituted C5-C7 aryl), optionally        substituted alkyl-aryl (e.g., an alkyl-aryl comprising at least        one of an optionally substituted C1-C6 alkyl, an optionally        substituted C5-C7 aryl, or combinations thereof), optionally        substituted heteroaryl, optionally substituted alkyl (e.g., a        C1-C6 linear or branched alkyl optionally substituted with one        or more halogen, cycloalkyl (e.g., a C3-C6 cycloalkyl), or aryl        (e.g., C5-C7 aryl)), optionally substituted alkoxyl group (e.g.,        a methoxy, ethoxy, butoxy, propoxy, pentoxy, or hexoxy; wherein        the alkoxyl may be substituted with one or more halogen, alkyl,        haloalky, fluoroalkyl, cycloalkyl (e.g., a C3-C6 cycloalkyl), or        aryl (e.g., C5-C7 aryl)), optionally substituted cycloalkyl,        optionally substituted heterocyclyl, —P(O)(OR′)R″, —P(O)R′R″,        OP(O)(OR′)R″, —OP(O)R′R″, —Cl, —F, —Br, —I, —CF₃, —CN,        —NR′SO₂NR′R″, —NR′CONR′R″, —CONR′COR″, —NR′C(═N—CN)NR′R″,        —C(═N—CN)NR′R″, —NR′C(═N—CN)R″, —NR′C(═C—NO₂)NR′R″, —SO₂NR′COR″,        —NO₂, —CO₂R′, —C(C═N—OR′)R″, —CR′═CR′R″, —CCR′,        —S(C═O)(C═N—R′)R″, —SF₅ and —OCF₃, wherein at least one W, X, Y,        Z, G, G′, R, R′, R″, Q1-Q4, or A is modified to be covalently        joined to a PTM, a chemical linking group (L), a ULM, CLM, or        combination thereof;    -   each of x, y, and z of Formulas (a1) through (e) are        independently 0, 1, 2, 3, 4, 5, or 6;    -   R′ and R″ of Formulas (a1) through (e) are independently        selected from a bond, H, optionally substituted linear or        branched alkyl, optionally substituted cycloalkyl, optionally        substituted aryl, optionally substituted heteroaryl, optionally        substituted heterocyclic, —C(═O)R, optionally substituted        heterocyclyl;    -   n′ of Formulas (a1) through (e) is an integer from 1-10 (e.g.        1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);    -   represents a single bond or a double bond; and    -   of Formulas (a1) through (e) represents a bond that may be        stereospecific ((R) or (S)) or non-stereospecific.

In any aspect or embodiment described herein, the CLM comprises achemical structure selected from the group consisting of:

wherein:

-   -   W of Formulas (a1) through (e) [e.g., (a1), (a2), (a3), (a4),        (b), (c), (d1), (d2), and/or (e)] is independently selected from        the group CH₂, O, CHR, C═O, SO₂, NH, N, optionally substituted        cyclopropyl group, optionally substituted cyclobutyl group, and        N-alkyl;    -   W₃ of Formulas (a1) through (e) is selected from C or N;    -   X of Formulas (a1) through (e) is independently selected from        the group O, S and CH₂;    -   Y of Formulas (a1) through (e) is independently selected from        the group CH₂, —C═CR′, NH, N-alkyl, N-aryl, N-hetaryl,        N-cycloalkyl, N-heterocyclyl, O, and S;    -   Z of Formulas (a1) through (e) is independently selected from        the group O, and S or CH2 except that both X and Z cannot be CH₂        or absent;    -   G and G′ of Formulas (a1) through (e) are independently selected        from the group H, optionally substituted linear or branched        alkyl, OH, R′OCOOR, R′OCONRR″, CH₂-heterocyclyl optionally        substituted with R′, and benzyl optionally substituted with R′;    -   Q1-Q4 of Formulas (a1) through (e) represent a carbon C or N        substituted with a group independently selected from H, R, N or        N-oxide;    -   A of Formulas (a1) through (e) is independently selected from        the group H, optionally substituted linear or branched alkyl,        cycloalkyl, Cl and F;    -   n of Formulas (a1) through (e) represent an integer from 1 to 10        (e.g., 1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);    -   R of Formulas (a1) through (e) comprises, but is not limited to:        a bond, H, —C(═O)R′ (e.g., a carboxy group), —CONR′R″ (e.g., an        amide group), —OR′ (e.g., OH), —NR′R″ (e.g. an amine group),        —SR′, —SO2R′, —SO2NR′R″, —CR′R″—, —CR′NR′R″—, (—CR′O)_(n)R″,        optionally substituted aryl (e.g., an optionally substituted        C5-C7 aryl), optionally substituted alkyl-aryl (e.g., an        alkyl-aryl comprising at least one of an optionally substituted        C1-C6 alkyl, an optionally substituted C5-C7 aryl, or        combinations thereof), optionally substituted hetaryl,        -optionally substituted linear or branched alkyl (e.g., a C1-C6        linear or branched alkyl optionally substituted with one or more        halogen, cycloalkyl (e.g., a C3-C6 cycloalkyl), or aryl (e.g.,        C5-C7 aryl)), optionally substituted alkoxyl group (e.g., a        methoxy, ethoxy, butoxy, propoxy, pentoxy, or hexoxy; wherein        the alkoxyl may be substituted with one or more halogen, alkyl,        haloalky, fluoroalkyl, cycloalkyl (e.g., a C3-C6 cycloalkyl), or        aryl (e.g., C5-C7 aryl)), optionally substituted cycloalkyl,        optionally substituted heterocyclyl, —P(O)(OR′)R″, —P(O)R′R″,        OP(O)(OR′)R″, —OP(O)R′R″, —Cl, —F, —Br, —I, —CF3, —CN,        —NR′SO2NR′R″, —NR′CONR′R″, —CONR′COR″, —NR′C(═N—CN)NR′R″,        —C(═N—CN)NR′R″, —NR′C(═N—CN)R″, —NR′C(═C—NO2)NR′R″, —SO2NR′COR″,        —NO2, —CO2R′, —C(C═N—OR′)R″, —CR′═CR′R″, —CCR′,        —S(C═O)(C═N—R′)R″, —SF5 and —OCF3, wherein at least one of W, X,        Y, Z, G, G′, R, R′, R″, Q1-Q4, or A is covalently joined        (directly or indirectly, e.g., via a functional group or an        atom, such as O, S, N) to a PTM, a chemical linking group (L), a        ULM, CLM, or combination thereof;    -   each of x, y, and z of Formulas (a1) through (e) are        independently 0, 1, 2, 3, 4, 5, or 6;    -   R′ and R″ of Formulas (a1) through (e) are independently        selected from a bond, H, optionally substituted linear or        branched alkyl, optionally substituted cycloalkyl, optionally        substituted aryl, optionally substituted heteroaryl, optionally        substituted heterocyclic, —C(═O)R, optionally substituted        heterocyclyl;    -   n′ of Formulas (a1) through (e) is an integer from 1-10 (e.g.,        1-4, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10); and    -   of Formulas (a1) through (e) represents a bond that may be        stereospecific ((R) or (S)) or non-stereospecific.

In any aspect or embodiment described herein, the CLM or ULM is selectedfrom the structure of Formula (g):

wherein:

-   -   W of Formula (g) is independently selected from the group CH₂,        O, C═O, NH, and N-alkyl;    -   A of Formula (g) is selected from a H, methyl, or optionally        substituted linear or branched alkyl;    -   n is an integer from 1 to 4;    -   R of Formula (g) is independently selected from a bond, H, O,        OH, N, NH, NH₂, Cl, —F, —Br, —I, methyl, optionally substituted        linear or branched alkyl (e.g., optionally substituted linear or        branched C1-C6 alkyl), optionally substituted linear or branched        alkoxy (e.g., optionally substituted linear or branched C1-C6        alkoxy), -alkyl-aryl (e.g., an -alkyl-aryl comprising at least        one of C1-C6 alkyl, C4-C7 aryl, or a combination thereof), aryl        (e.g., C5-C7 aryl), amine, amide, or carboxy), wherein at least        one R or W is modified to be covalently joined to a PTM, a        chemical linking group (L), a ULM, CLM, or combination thereof;        and    -   of Formula (g) represents a bond that may be stereospecific ((R)        or (S)) or non-stereo specific.

In any aspect or embodiment described herein, the CLM or ULM is selectedfrom the group consisting of:

wherein:

-   -   W is C═O or CH₂;    -   N* is a nitrogen atom that is covalently linked to the PTM or        linker, or that is shared with the PTM or linker (L) (e.g., a        heteroatom shared with an optionally substituted heterocyclyl of        the linker (L) or PTM); and    -   indicates the point of attachment of the CLM or ULM to the        linker (L) or PTM.

In any aspect or embodiment described herein, R is selected from: H, O,OH, N, NH, NH₂, C1-C6 alkyl, C1-C6 alkoxy, -alkyl-aryl (e.g., an-alkyl-aryl comprising at least one of C1-C6 alkyl, C4-C7 aryl, or acombination thereof), aryl (e.g., C5-C7 aryl), amine, amide, orcarboxy).

In any aspect or embodiment described herein, at least one R (e.g. an Rgroup selected from the following H, O, OH, N, NH, NH₂, C1-C6 alkyl,C1-C6 alkoxy, -alkyl-aryl (e.g., an -alkyl-aryl comprising at least oneof C1-C6 alkyl, C4-C7 aryl, or a combination thereof), aryl (e.g., C5-C7aryl), amine, amide, or carboxy) or W is modified to be covalentlyjoined to a PTM, a chemical linker group (L), a ULM, a CLM, or acombination thereof

In any aspect or embodiment described herein, the W, X, Y, Z, G, G′, R,R′, R″, Q1-Q4, and A of Formulas (a) through (g) can independently becovalently coupled to a linker and/or a linker to which is attached oneor more PTM, ULM, or CLM groups.

In any of the aspects or embodiments described herein, n is an integerfrom 1 to 4, and each R is independently selected functional groups oratoms, for example, O, OH, N, —Cl, —F, C1-C6 alkyl, C1-C6 alkoxy,-alkyl-aryl (e.g., an -alkyl-aryl comprising at least one of C1-C6alkyl, C4-C7 aryl, or a combination thereof), aryl (e.g., C5-C7 aryl),amine, amide, or carboxy, on the aryl or heteroaryl of the CLM, andoptionally, one of which is modified to be covalently joined to a PTM, achemical linker group (L), a ULM, CLM or combination thereof.

More specifically, non-limiting examples of CLMs include those shownbelow as well as those “hybrid” molecules that arise from thecombination of one or more of the different features shown in themolecules below wherein at least one R or W is modified to be covalentlyjoined to a PTM, a chemical linking group (L), a ULM, CLM, orcombination thereof.

In any aspect or embodiment described herein, the CLM comprises achemical structure selected from the group:

wherein:

-   -   W is independently selected from CH₂, O, CHR, C═O, SO₂, NH, N,        optionally substituted cyclopropyl group, optionally substituted        cyclobutyl group, and N-alkyl (e.g., CH₂, CHR, C═O, SO₂, NH, and        N-alkyl);    -   Q₁, Q₂, Q₃, Q₄, Q₅ are each independently represent a carbon C        or N substituted with a group independently selected from R′, N        or N-oxide;    -   R¹ is selected from absent (i.e., a bond), H, OH, CN, C1-C3        alkyl, C═O;    -   R² is selected from the group absent (i.e., a bond), H, OH, CN,        C1-C3 alkyl, CHF₂, CF₃, CHO, C(═O)NH₂;    -   R³ is selected from a bond, H, alkyl (e.g., C1-C6 or C1-C3        alkyl), substituted alkyl (e.g., substituted C1-C6 or C1-C3        alkyl), alkoxy (e.g., C1-C6 or C1-C3 alkoxyl), substituted        alkoxy (e.g., substituted C1-C6 or C1-C3 alkoxyl);    -   R⁴ is selected from a bond, H, alkyl, substituted alkyl;    -   R⁵ and R⁶ are each independently a bond, H, halogen, C(═O)R′,        CN, OH, CF₃;    -   X is C, CH, C═O, or N;    -   X₁ is C═O, N, CH, or CH₂;    -   R′ is selected from a bond, H, halogen, amine, alkyl (e.g.,        C1-C3 alkyl), substituted alkyl (e.g., substituted C1-C3 alkyl),        alkoxy (e.g., C1-C3 alkoxyl), substituted alkoxy (e.g.,        substituted C1-C3 alkoxyl), NR²R³, C(═O)OR², optionally        substituted phenyl;    -   n is 0-4;    -   is a single or double bond; and    -   the CLM is covalently joined to a PTM, a chemical linker group        (L), a ULM, CLM or combination thereof.

In any aspect or embodiment described herein, the CLM is covalentlyjoined to a PTM or a chemical linker group (L) via an R group (such as,R, R¹, R², R³, R⁴ or R′), W, X, or a Q group (such as, Q₁, Q₂, Q₃, Q₄,or Q₅).

In any aspect or embodiment described herein, the CLM is covalentlyjoined to a PTM or a chemical linker group (L) via W, X, R, R¹, R², R³,R⁴, R⁵, R′, Q₁, Q₂, Q₃, Q₄, and Q₅.

In any aspect or embodiment described herein, the W, X, R¹, R², R³, R⁴,R′, Q₁, Q₂, Q₃, Q₄, and Q₅ can independently be covalently coupled to alinker and/or a linker to which is attached to one or more PTM, ULM, CLMgroups.

More specifically, non-limiting examples of CLMs include those shownbelow as well as “hybrid” molecules or compounds that arise fromcombining one or more features of the following compounds:

wherein:

-   -   W is independently selected from the group CH₂, CHR, C═O, SO₂,        NH, and N-alkyl;    -   R¹ is selected from the group absent (i.e., a bond), H, CH, CN,        C1-C3 alkyl;    -   R² is selected from a bond, H or a C1-C3 alkyl;    -   R³ is selected from a bond, H, alkyl, substituted alkyl, alkoxy,        substituted alkoxy;    -   R⁴ is selected from a bond, methyl or ethyl;    -   R⁵ is selected from a bond, H or halo;    -   R⁶ is selected from a bond, H or halo;    -   n is an integer from 0-4;    -   R and R′ are independently a bond, H, a functional group, or an        atom (e.g., H, halogen (e.g., —Cl, —F), amine, C1-C3 alkyl,        C1-C3 alkyl, C1-C3 alkoxyl, NR²R³, or C(═O)OR²); or an        attachment point for a PTM or a chemical linker group (L);    -   Q₁ and Q₂ are each independently C or N substituted with a group        independently selected from H or C1-C3 alkyl;    -   is a single or double bond.

In any aspect or embodiment described herein, the W, R¹, R², Q₁, Q₂, Q₃,Q₄, R, and R′ can independently be covalently coupled to a linker and/ora linker to which is attached one or more PTM groups.

In any aspect or embodiment described herein, the R¹, R², Q₁, Q₂, Q₃,Q₄, R, and R′ can independently be covalently coupled to a linker and/ora linker to which is attached one or more PTM groups.

In any aspect or embodiment described herein, the Q₁, Q₂, Q₃, Q₄, R, andR′ can independently be covalently coupled to a linker and/or a linkerto which is attached one or more PTM groups.

In any aspect or embodiment described herein, R is modified to becovalently joined to the linker group (L), or a PTM, or combinationthereof.

In any aspect or embodiment described herein, the CLM is selected from:

wherein R′ is a halogen and R¹ is as described herein.

In any aspect or embodiment described herein, “CLM” can be an imide thatbinds to cereblon E3 ligase. These imides and linker attachment pointcan be, but not be limited to one of the following structures:

In any aspect or embodiment described herein, the ULM is selected fromthe group consisting of:

wherein:

-   -   of the CLM indicates the point of attachment with a linker group        or a PTM; and    -   N* is a nitrogen atom that is shared with the chemical linker        group or PTM

In any aspect or embodiment described herein, the ULM is selected fromthe group consisting of:

wherein:

-   -   of the ULM indicates the point of attachment with a linker group        or a PTM;    -   N* is a nitrogen atom that is shared with the chemical linker        group or PTM; and    -   W, Q4, and Q5 are each defined as described in any aspect or        embodiment described herein.

Exemplary Tankers

In any aspect or embodiment described herein, the compounds as describedherein include a PTM chemically linked to a ULM (e.g., CLM) via achemical linker (L). In certain embodiments, the linker group Lcomprises one or more covalently connected structural units (e.g.,-A^(L) _(1 . . .) (A^(L))_(q)- or -(A^(L))_(q)-), wherein A^(L) ₁ is agroup coupled to PTM, and (A^(L))_(q) is a group coupled to ULM.

In any aspect or embodiment described herein, the linker (L) to a ULM(e.g., CLM) connection is a stable L-ULM connection. For example, incertain embodiments, when a linker (L) and a ULM are connected via aheteroatom (e.g., N, O, S), any additional heteroatom, if present, isseparated by at least a carbon atom (e.g., —CH₂—), such as with anacetal or aminal group. By way of further example, in certainembodiments described herein, when a linker (L) and a ULM are connectedvia a heteroatom, the heteroatom is not part of an ester.

In any aspect or embodiment described herein, the linker group L is abond or a chemical linker group represented by the formula-(A^(L))_(q)-, wherein A is a chemical moiety and q is an integer from1-100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53,54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,72, 73, 74, 75, 76, 77, 78, 79, or 80), and wherein L is covalentlybound to both the PTM and the ULM, and provides for binding of the PTMto the protein target and the ULM to an E3 ubiquitin ligase toeffectuate target protein ubiquitination.

In any aspect or embodiment described herein, the linker group L is abond or a chemical linker group represented by the formula-(A^(L))_(q)-, wherein A is a chemical moiety and q is an integer from6-30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, or 25), and wherein L is covalently bound toboth the PTM and the ULM, and provides for binding of the PTM to theprotein target and the ULM to an E3 ubiquitin ligase in sufficientproximity to result in target protein ubiquitination.

In any aspect or embodiment described herein, the linker group L is-(A^(L))_(q)-, wherein:

-   -   (A^(L))_(q) is a group which connects a ULM (e.g., CLM), to PTM;    -   q of the linker is an integer greater than or equal to 1;    -   each A^(L) is independently selected from the group consisting        of, a bond, CR^(L1)R^(L2), O, S, SO, SO₂, NR^(L3), SO₂NR^(L3),        SONR^(L3), CONR^(L3), NR^(L3)CONR^(L4), NR^(L3)SO₂NR^(L4), CO,        CR^(L1)═CR^(L2), C≡C, SiR^(L1)R^(L2), P(O)R^(L1), P(O)OR^(L1),        NR^(L3)C(═NCN)NR^(L4), NR^(L3)C(═NCN), NR^(L3)C(═CNO₂)NR^(L4),        C₃₋₁₁cycloalkyl optionally substituted with 1-6 R^(L1) and/or        R^(L2) groups, C₅₋₁₃ spirocycloalkyl optionally substituted with        1-9 R^(L1) and/or R^(L2) groups, C₃₋₁₁heterocyclyl optionally        substituted with 1-6 R^(L1) and/or R^(L2) groups, C₅₋₁₃        spiroheterocyclyl optionally substituted with 1-8 R^(L1) and/or        R^(L2) groups, aryl optionally substituted with 1-6 R^(L1)        and/or R^(L2) groups, heteroaryl optionally substituted with 1-6        R^(L1) and/or R^(L2) groups, where R^(L1) or R^(L2), each        independently are optionally linked to other groups to form        cycloalkyl and/or heterocyclyl moiety, optionally substituted        with 1-4 R¹⁵ groups; and    -   R^(L1), R^(L2), R^(L3), R^(L4) and R^(L5) are, each        independently, H, halo, C₁₋₈alkyl, OC₁₋₈alkyl, SC₁₋₈alkyl,        NHC₁₋₈alkyl, N(C₁₋₈alkyl)₂, C₃₋₁₁cycloalkyl, aryl, heteroaryl,        C₃₋₁₁heterocyclyl, OC₃₋₈cycloalkyl, SC₃₋₈cycloalkyl,        NHC₃₋₈cycloalkyl, N(C₃₋₅cycloalkyl)₂,        N(C₃₋₈cycloalkyl)(C₁₋₈alkyl), OH, NH₂, SH, SO₂C₁₋₈alkyl,        P(O)(OC₁₋₈alkyl)(C₁₋₈alkyl), P(O)(OC₁₋₈alkyl)₂, CC—C₁₋₈alkyl,        CCH, CH═CH(C₁₋₈alkyl), C(C₁₋₈alkyl)═CH(C₁₋₈alkyl),        C(C₁₋₈alkyl)═C(C₁₋₈alkyl)₂, Si(OH)₃, Si(C₁₋₈alkyl)₃,        Si(OH)(C₁₋₈alkyl)₂, COC₁₋₈alkyl, CO₂H, halogen, CN, CF₃, CHF₂,        CH₂F, NO₂, SF₅, SO₂NHC₁₋₈alkyl, SO₂N(C₁₋₈alkyl)₂, SONHC₁₋₈alkyl,        SON(C₁₋₈alkyl)₂, CONHC₁₋₈alkyl, CON(C₁₋₈alkyl)₂,        N(C₁₋₈alkyl)CONH(C₁₋₈alkyl), N(C₁₋₈alkyl)CON(C₁₋₈alkyl)₂,        NHCONH(C₁₋₈alkyl), NHCON(C₁₋₈alkyl)₂, NHCONH₂,        N(C₁₋₈alkyl)SO₂NH(C₁₋₈alkyl), N(C₁₋₈alkyl) SO₂N(C₁₋₈alkyl)₂, NH        SO₂NH(C₁₋₈alkyl), NH SO₂N(C₁₋₈alkyl)₂, NH SO₂NH₂.

In certain embodiments, q is an integer greater than or equal to 1.

In certain embodiments, e.g., where q of the linker is greater than 2,(A^(L))_(q) is a group which is A^(L) ₁ and (A^(L))_(q) wherein thelinker couples a PTM to a ULM.

In certain embodiments, e.g., where q of the linker is 2, A^(L) ₂ is agroup which is connected to A^(L) ₁ and to a ULM.

In certain embodiments, e.g., where q of the linker is 1, the structureof the linker group L is -A^(L) ₁-, and A^(L) ₁ is a group whichconnects a ULM moiety to a PTM moiety.

In any aspect or embodiment described herein, the unit A^(L) of linker(L) comprises a group represented by a general structure selected fromthe group consisting of:

-   -   —NR(CH₂)_(n)-(lower alkyl)-, —NR(CH₂)_(n)-(lower alkoxyl)-,        —NR(CH₂)_(n)-(lower alkoxyl)-OCH₂—, —NR(CH₂)_(n)-(lower        alkoxyl)-(lower alkyl)-OCH₂—, —NR(CH₂)_(n)-(cycloalkyl)-(lower        alkyl)-OCH₂—, —NR(CH₂)_(n)-(heterocycloalkyl)-,        —NR(CH₂CH₂O)_(n)-(lower alkyl)-O—CH₂—,        —NR(CH₂CH₂O)_(n)-(heterocycloalkyl)-O—CH₂—,        —NR(CH₂CH₂O)_(n)-Aryl-O—CH₂—,        NR(CH₂CH₂O)_(n)-(heteroaryl)-O—CH₂—, —NR(CH₂CH₂O)_(n)-(cyclo        alkyl)-O-(heteroaryl)-O—CH₂—, —NR(CH₂CH₂O)_(n)-(cyclo        alkyl)-O-Aryl-O—CH₂—, —NR(CH₂CH₂O)_(n)-(lower        alkyl)-NH-Aryl-O—CH₂—, —NR(CH₂CH₂O)_(n)-(lower        alkyl)-O-Aryl-CH₂, —NR(CH₂CH₂O)_(n)-cycloalkyl-O-Aryl-,        —NR(CH₂CH₂O)_(n)-cycloalkyl-O-(heteroaryl)l-,        —NR(CH₂CH₂)_(n)-(cycloalkyl)-O-(heterocyclyl)-CH₂,        —NR(CH₂CH₂)_(n)-(heterocyclyl)-(heterocyclyl)-CH₂, and        —N(R1R2)-(heterocyclyl)-CH₂; where    -   n of the linker can be 0 to 10;    -   R of the linker can be H, or lower alkyl; and    -   R1 and R2 of the linker can form a ring with the connecting N.

In any aspect or embodiment described herein, the linker (L) includes anoptionally substituted C₁-C₅₀alkyl (e.g., C₁, C₂, C₃, C₄, C₅, C₆, C₇,C₈, C₉, C₁₀, C₁₁, C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉, C₂₀, C₂₁, C₂₂,C₂₃, C₂₄, C₂₅, C₂₆, C₂₇, C₂₈, C₂₉, C₃₀, C₃₁, C₃₂, C₃₃, C₃₄, C₃₅, C₃₆,C₃₇, C₃₈, C₃₉, C₄₀, C₄₁, C₄₂, C₄₃, Cn, C₄₅, C₄₆, C₄₇, C₄₈, C₄₉, or C₅₀alkyl, and including all implied subranges, e.g., C1-C10, C1-C20;C2-C10, C2-20; C10-C20, C10-C50 etc.), wherein each carbon is optionallyindependently substituted or replaced with (1) a heteroatom selectedfrom N, O, S, P, or Si atoms that has an appropriate number ofhydrogens, substitutions, or both to complete valency, (2) an optionallysubstituted cycloalkyl or bicyclic cycloalkyl, (3) an optionallysubstituted heterocyloalkyl or bicyclic heterocyloalkyl, (4) anoptionally substituted aryl or bicyclic aryl, or (5) optionallysubstituted heteroaryl or bicyclic heteroaryl. In any aspect orembodiment described herein, the linker (L) does not haveheteroatom-heteroatom bonding (e.g., no heteroatoms are covalentlylinked or adjacently located).

In any aspect or embodiment described herein, the linker (L) includes anoptionally substituted C₁-C₅₀ alkyl (e.g., C₁, C₂, C₃, C₄, C₅, C₆, C₇,C₈, C₉, C₁₀, C₁₁, C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇, C₁₈, C₁₉, C₂₀, C₂₁, C₂₂,C₂₃, C₂₄, C₂₅, C₂₆, C₂₇, C₂₈, C₂₉, C₃₀, C₃₁, C₃₂, C₃₃, C₃₄, C₃₅, C₃₆,C₃₇, C₃₈, C₃₉, C₄₀, C₄₁, C₄₂, C₄₃, C₄₄, C₄₅, C₄₆, C₄₇, C₄₈, C₄₉, or C₅₀alkyl), wherein:

-   -   each carbon is optionally independently substituted or replaced        with CR^(L1)R^(L2), O, S, SO, SO₂, NR^(L3), SO₂NR^(L3),        SONR^(L3), CONR^(L3), NR^(L3)CONR^(L4), NR^(L3)SO₂NR^(L4), CO,        CR^(L1)═CR^(L2), C≡C, SiR^(L1)R^(L2), P(O)R^(L1), P(O)OR^(L1),        NR^(L3)C(═NCN)NR^(L4), NR^(L3)C(═NCN), NR^(L3)C(═CNO₂)NR^(L4),        C₃₋₁₁cycloalkyl optionally substituted with 1-6 R^(L1) and/or        R^(L2) groups, C₅₋₁₃ spirocycloalkyl optionally substituted with        1-9 R^(L1) and/or R^(L2) groups, C₃₋₁₁ heterocyclyl optionally        substituted with 1-6 R^(L1) and/or R^(L2) groups, C₅₋₁₃        spiroheterocyclyl optionally substituted with 1-8 R^(L1) and/or        R^(L2) groups, aryl optionally substituted with 1-6 R^(L1)        and/or R^(L2) groups, heteroaryl optionally substituted with 1-6        R^(L1) and/or R^(L2) groups, where R^(L1) or R^(L2), each        independently are optionally linked to other groups to form a        cycloalkyl and/or a heterocyclyl moiety, optionally substituted        with 1-4 R^(L5) groups; and    -   R^(L1), R^(L2), R^(L3), R^(L4) and R^(L5) are, each        independently, H, halo, C₁₋₈alkyl, OC₁₋₈alkyl, SC₁₋₈alkyl,        NHC₁₋₈alkyl, N(C₁₋₅alkyl)₂, C₃₋₁₁cycloalkyl, aryl, heteroaryl,        C₃₋₁₁heterocyclyl, OC₃₋₈cycloalkyl, SC₃₋₈cycloalkyl,        NHC₃₋₈cycloalkyl, N(C₃₋₅cycloalkyl)₂,        N(C₃₋₈cycloalkyl)(C₁₋₈alkyl), OH, NH₂, SH, SO₂C₁₋₈alkyl,        P(O)(OC₁₋₈alkyl)(C₁₋₈alkyl), P(O)(OC₁₋₈alkyl)₂, CC—C₁₋₈alkyl,        CCH, CH═CH(C₁₋₈alkyl), C(C₁₋₈alkyl)═CH(C₁₋₈alkyl),        C(C₁₋₈alkyl)═C(C₁₋₈alkyl)₂, Si(OH)₃, Si(C₁₋₈alkyl)₃,        Si(OH)(C₁₋₈alkyl)₂, COC₁₋₈alkyl, CO₂H, halogen, CN, CF₃, CHF₂,        CH₂F, NO₂, SF₅, SO₂NHC₁₋₈alkyl, SO₂N(C₁₋₈alkyl)₂, SONHC₁₋₈alkyl,        SON(C₁₋₈alkyl)₂, CONHC₁₋₈alkyl, CON(C₁₋₈alkyl)₂,        N(C₁₋₈alkyl)CONH(C₁₋₈alkyl), N(C₁₋₈alkyl)CON(C₁₋₈alkyl)₂,        NHCONH(C₁₋₈alkyl), NHCON(C₁₋₈alkyl)₂, NHCONH₂,        N(C₁₋₈alkyl)SO₂NH(C₁₋₈alkyl), N(C₁₋₈alkyl) SO₂N(C₁₋₈alkyl)₂, NH        SO₂NH(C₁₋₈alkyl), NH SO₂N(C₁₋₈alkyl)₂, NH SO₂NH₂.

In any aspect or embodiment described herein, the linker group isoptionally substituted an optionally substituted C₁-C₅₀alkyl (e.g., C₁,C₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁, C₁₂, C₁₃, C₁₄, C₁₅, C₁₆, C₁₇,C₁₈, C₁₉, C₂₀, C₂₁, C₂₂, C₂₃, C₂₄, C₂₅, C₂₆, C₂₇, C₂₈, C₂₉, C₃₀, C₃₁,C₃₂, C₃₃, C₃₄, C₃₅, C₃₆, C₃₇, C₃₈, C₃₉, C₄₀, C₄₁, C₄₂, C₄₃, C₄₄, C₄₅,C₄₆, C₄₇, C₄₈, C₄₉, or C₅₀ alkyl, and including all implied subranges,e.g., C1-C10, C1-C20; C2-C10, C2-20; C10-C20, C10-C50 etc.), whereineach carbon atom optionally substituted or replaced with: a O, N, S, Por Si atom that has an appropriate number of hydrogens, substitutions(e.g., OH, halo, alkyl, methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy,methoxy, etc.), or both to complete valency; an optionally substitutedaryl (e.g., an optionally substituted C5 or C6 aryl) or bicyclic aryl(e.g, an optionally substituted C5-C20 bicyclic heteroaryl); anoptionally substituted heteroaryl (e.g., an optionally substituted C5 orC6 heteroaryl) or bicyclic heteroaryl (e.g., an optionally substitutedheteroaryl or bicyclic heteroaryl having one or more heteroatomsselected from N, O, S, P, and Si that has an appropriate number ofhydrogens, substitutions (e.g., OH, halo, alkyl, methyl, ethyl,haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or both to completevalency); an optionally substituted C1-C6 alkyl; an optionallysubstituted C1-C6 alkenyl; an optionally substituted C1-C6 alkynyl; anoptionally substituted cycloalkyl (e.g., an optionally substituted C3-C7cycloalkyl) or bicyclic cycloalkyl (e.g., an optionally substitutedC5-C20 bicyclic cycloalkyl); or an optionally substitutedheterocycloalkyl (e.g., an optionally substituted 3-, 4-, 5-, 6-, or7-membered heterocyclic group) or bicyclicheteroalkyl (e.g., anoptionally substituted heterocyclo alkyl bicyclicheteroalkyl having oneor more heteroatoms selected from N, O, S, P, or Si atoms that has anappropriate number of hydrogens, substitutions (e.g., OH, halo, alkyl,methyl, ethyl, haloalkyl, hydroxyalkyl, alkoxy, methoxy, etc.), or bothto complete valency). In any aspect or embodiment described herein, theoptionally substituted alkyl linker is optionally substituted with oneor more OH, halo, linear or branched C1-C6 alkyl (such as methyl orethyl), linear or branched C1-C6 haloalkyl, linear or branched C1-C6hydroxyalkyl, or linear or branched C1-C6 alkoxy (e.g., methoxy).

In any aspect or embodiment described herein, the linker (L) does nothave heteroatom-heteroatom bonding (e.g., no heteroatoms are covalentlylinked or adjacently located).

In any aspect or embodiment described herein, the linker (L) includesabout 1 to about 50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,or 50) alkylene glycol units that are optionally substituted, whereincarbon or oxygen may be substituted with a heteroatom selected from N,S, P, or Si atoms with an appropriate number of hydrogens to completevalency.

In any aspect or embodiment described herein, the L is selected from thegroup consisting of:

wherein:

-   -   N* is a nitrogen atom that is covalently linked to the CLM or        PTM, or that is shared with the CLM or PTM;    -   each m, n, o, and p is independently 0, 1, 2, 3, 4, 5, 6, 7, 8,        9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; and    -   the chemical linker group is optionally substituted with 1, 2,        3, or 4 substitutions independently selected from a halogen        (e.g., F or Cl) and a C₁₋₄ alkyl

In any aspect or embodiment described herein, the until A^(L) of thelinker (L) comprises a structure selected from the group consisting of:

wherein N* is a nitrogen atom that is covalently linked to the ULM orPTM, or that is shared with the ULM or PTM.

In any aspect or embodiment described herein, the until A^(L) of thelinker (L) comprises a structure selected from the group consisting of:

wherein N* is a nitrogen atom that is covalently linked to the ULM orPTM, or that is shared with the ULM or PTM.

In any aspect or embodiment described herein, the unit A^(L) of thelinker (L) comprises a structure selected from the group consisting of:

wherein:

-   -   N* is a nitrogen atom that is covalently linked to the ULM or        PTM, or that is shared with the ULM or PTM; and    -   each m, n, o, p, q, and r is independently 0, 1, 2, 3, 4, 5, 6,        7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

In any aspect or embodiment described herein, the unit A^(L) of thelinker (L) is selected from:

wherein dashed line or * indicates the site that is covalently linked tothe ULM or PTM, or that is shared with the ULM or PTM.

In any aspect or embodiment described herein, the unit A^(L) of thelinker (L) is selected from:

wherein dashed line or * indicates the site that is covalently linked tothe CLM or the PTM, or that is shared with the CLM or the PTM.

In any aspect or embodiment described herein, the unit A^(L) of linker(L) comprises a group represented by a general structure selected fromthe group consisting of:

whereinm, n, o, p, q, and r of the linker are independently 0, 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20;when m, n, o, p, q, and r are zero, N—O or O—O bond is absent,X of the linker is H or F

where each n and m of the linker can independently be 0, 1, 2, 3, 4, 5,or 6.

In any aspect or embodiment described herein, the unit A^(L) of linker(L) is selected from the group consisting of:

wherein each m and n is independently selected from 0, 1, 2, 3, 4, 5, or6.

In any aspect or embodiment described herein, the unit A^(L) of linker(L) is selected from the group consisting of:

wherein each m, n, o, p, q, r, and s is independently 0, 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

In any aspect or embodiment described herein, the unit A^(L) of linker(L) is selected from the group consisting of:

In any aspect or embodiment described herein, the linker (L) comprises astructure selected from the structure shown below:

wherein:

-   -   W^(L1) and W^(L2) are each independently absent, a 4-8 membered        ring with 0-4 heteroatoms, optionally substituted with R^(Q),        each R^(Q) is independently a H, halo, OH, CN, CF₃, optionally        substituted linear or branched C₁-C₆ alkyl, optionally        substituted linear or branched C₁-C₆ alkoxy, or 2 R^(Q) groups        taken together with the atom they are attached to, form a 4-8        membered ring system containing 0-4 heteroatoms;    -   Y^(L1) is each independently a bond, optionally substituted        linear or branched C₁-C₆ alkyl and optionally one or more C        atoms are replaced with O or NR^(YL1), optionally substituted        C₁-C₆ alkene and optionally one or more C atoms are replaced        with O, optionally substituted C₁-C₆ alkyne, and optionally one        or more C atoms are replaced with O, or optionally substituted        linear or branched C₁-C₆ alkoxy;    -   R^(YL1) is H, or optionally substituted linear or branched C₁₋₆        alkyl;    -   n is 0-10; and    -   and        indicates the attachment point to the PTM or ULM moieties.

In any aspect or embodiment described herein, the linker (L) comprises astructure selected from the structure shown below:

wherein:

-   -   W^(L1) and W^(L2) are each independently absent, piperazine,        piperidine, morpholine, optionally substituted with R^(Q), each        R^(Q) is independently a H, —Cl—, —F—, OH, CN, CF₃, optionally        substituted linear or branched C₁-C₆ alkyl (e.g. methyl, ethyl),        optionally substituted linear or branched C₁-C₆ alkoxy (e.g.        methoxy, ethoxy);    -   Y^(L1) is each independently a bond, optionally substituted        linear or branched C₁-C₆ alkyl and optionally one or more C        atoms are replaced with O or NR^(YL1); optionally substituted        C₁-C₆ alkene and optionally one or more C atoms are replaced        with O, optionally substituted C₁-C₆ alkyne and optionally one        or more C atoms are replaced with O, or optionally substituted        linear or branched C₁-C₆ alkoxy;    -   R^(YL1) is H, or optionally substituted linear or branched C₁₋₆        alkyl (e.g. methyl, ethyl);    -   n is 0-10; and    -   and        indicates the attachment point to the PTM or ULM moieties.

In any aspect or embodiment described herein, the linker (L) comprises astructure selected from the structure shown below:

wherein:

-   -   W^(L1) and W^(L2) are each independently absent, aryl,        heteroaryl, cyclic, heterocyclic, C₁₋₆ alkyl and optionally one        or more C atoms are replaced with O or NR^(YL1), C₁₋₆ alkene and        optionally one or more C atoms are replaced with O, C₁₋₆ alkyne        and optionally one or more C atoms are replaced with O,        bicyclic, biaryl, biheteroaryl, or biheterocyclic, each        optionally substituted with R^(Q), each R^(Q) is independently a        H, halo, OH, CN, CF₃, hydroxyl, nitro, C≡CH, C₂₋₆ alkenyl, C₂₋₆        alkynyl, optionally substituted linear or branched C₁-C₆ alkyl,        optionally substituted linear or branched C₁-C₆ alkoxy,        optionally substituted OC₁₋₃alkyl (e.g., optionally substituted        by 1 or more —F), OH, NH₂, NR^(Y1)R^(Y2), CN, or 2 R^(Q) groups        taken together with the atom they are attached to, form a 4-8        membered ring system containing 0-4 heteroatoms;    -   Y^(L1) is each independently a bond, NR^(YL1), O, S, NR^(YL2),        CR^(YL1)R^(YL2), C═O, C═S, SO, SO₂, optionally substituted        linear or branched C₁-C₆ alkyl and optionally one or more C        atoms are replaced with O; optionally substituted linear or        branched C₁-C₆ alkoxy;    -   Q^(L) is a 3-6 membered alicyclic, bicyclic, or aromatic ring        with 0-4 heteroatoms, optionally bridged, optionally substituted        with 0-6 R^(Q), each R^(Q) is independently H, optionally        substitute linear or branched C₁₋₆ alkyl (e.g., optionally        substituted by 1 or more halo, C₁₋₆ alkoxyl), or 2 R^(Q) groups        taken together with the atom they are attached to, form a 3-8        membered ring system containing 0-2 heteroatoms;    -   R^(YL1), R^(YL2) are each independently H, OH, optionally        substituted linear or branched C₁₋₆ alkyl (e.g., optionally        substituted by 1 or more halo, C₁₋₆ alkoxyl), or R¹, R² together        with the atom they are attached to, form a 3-8 membered ring        system containing 0-2 heteroatoms;    -   n is 0-10; and    -   and        indicates the attachment point to the PTM or ULM moieties.

In any aspect or embodiment described herein, the linker (L) comprises astructure selected from the structure shown below:

wherein:

-   -   W^(L1) and W^(L2) are each independently absent, cyclohexane,        cyclopentane, piperazine, piperidine, morpholine, C₁₋₆ alkyl and        optionally one or more C atoms are replaced with O or NR^(YL1),        C₁₋₆ alkene and optionally one or more C atoms are replaced with        O, or C₁₋₆ alkyne and optionally one or more C atoms are        replaced with O, each optionally substituted with R^(q), each        R^(Q) is independently a H, Cl—, —F—, OH, CN, CF₃, hydroxyl,        optionally substituted linear or branched C₁-C₆ alkyl (e.g.        methyl, ethyl), optionally substituted linear or branched C₁-C₆        alkoxy;    -   Y^(L1) is each independently a bond, NR^(YL1), O,        CR^(YL1)R^(YL2), C═O, optionally substituted linear or branched        C₁-C₆ alkyl and optionally one or more C atoms are replaced with        O or NR^(YL1), C₁₋₆ alkene and optionally one or more C atoms        are replaced with O, C₁₋₆ alkyne and optionally one or more C        atoms are replaced with O, or optionally substituted linear or        branched C₁-C₆ alkoxy;    -   Q^(L) is a 3-6 membered heterocyclic, heterobicyclic, or        heteroaryl ring, optionally substituted with 0-6 R^(q), each        R^(Q) is independently H, or optionally substituted linear or        branched C₁₋₆ alkyl (e.g., methyl or ethyl, optionally        substituted by 1 or more halo, C₁₋₆ alkoxyl);    -   R^(YL1), R^(YL2) are each independently H, optionally        substituted linear or branched C₁₋₆ alkyl (e.g., optionally        substituted by 1 or more halo, C₁₋₆ alkoxyl);    -   n is 0-10; and    -   and        indicates the attachment point to the PTM or ULM moieties.

Exemplary PTMs

The term “protein target moiety” or PTM is used to describe a smallmolecule which binds to LRRK2, and can be used to target the PTM forubiquitination and degradation. The compositions described belowexemplify members of LRRK2 binding moieties that can be used accordingto the present invention. These binding moieties are linked to theubiquitin ligase binding moiety preferably through a chemical linkinggroup in order to present the LRRK2 protein in proximity to theubiquitin ligase for ubiquitination and subsequent degradation.

In certain contexts, the term “target protein” is used to refer to theLRRK2 protein, a member of the ROCO protein family, that serves as anupstream central integrator of multiple signaling pathways that arecrucial for proper neuronal functioning, which is a target protein to beubiquitinated and degraded.

The compositions described herein exemplify the use of some of themembers of these types of small molecule target protein bindingmoieties.

In any aspect or embodiment described herein, the PTM is a smallmolecule that binds LRRK2. For example, in any aspect or embodimentdescribed herein, the PTM is represented by the chemical structurePTM-I:

wherein:

-   X is CH or N;-   W is O or S;-   Q is selected from the group:

wherein:

-   -   A¹, A², and A³ are each independently selected from N and CR⁶,        wherein for (a) no more than two of A¹, A², and A³ are        simultaneously N, ring B is an optionally substituted C3-C8        aryl, optionally substituted C3-C8 heteroaryl, optionally        substituted C3-C8 cycloalkyl, optionally substituted C3-C8        heterocycloalkyl, optionally substituted C3-C8 aryl, optionally        substituted C3-C8 aryl,    -   the        of Q indicates the point of attachment with the nitrogen, and    -   the        of Q indicates an optional covalent attachment to R_(PTM);

-   R² is H or C₁₋₄ alkyl;

-   R^(3A) and R^(3B) are each independently selected from H, halo, C₁₋₆    alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀    cycloalkyl, 5-14 membered heteroaryl, 4-14 membered    heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl,    5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered    heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a1), SR^(a1), C(O)R^(b1),    C(O)NR^(c1)R^(d1), C(O)OR^(a1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1),    NR^(c1)R^(d1), NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1),    NR^(c1)C(O)NR^(c1)R^(d1), C(═NR^(e1))R^(b1),    C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)(═NR^(e1))NR^(c1)R^(d1),    NR^(c1)S(O)R^(b1), NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1),    S(O)R^(b1), S(O)NR^(c1)R^(d1), S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);    wherein said C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl,    C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl, 5-14 membered heteroaryl, 4-14    membered heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇    cycloalkyl-C₁₋₄ alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and 4-10    membered heterocycloalkyl-C₁₋₄ alkyl of R¹ are each optionally    substituted with 1, 2, 3, 4, or 5 substituents independently    selected from Cy², Cy²-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl,    C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a1), SR^(a1), C(O)R^(b1),    C(O)NR^(c1)R^(d1), C(O)OR^(a1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1),    NR^(c1)R^(d1), NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1),    NR^(c1)C(O)NR^(c1)R^(d1), C(═NR^(e1))R^(b1),    C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)C(═NR^(e1))NR^(c1)R^(d1),    NR^(c1)S(O)R^(b1), NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1),    S(O)R^(b1), S(O)NR^(c1)R^(d1), S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);

-   or R^(3A) and R^(3B) together form a C₃₋₇ cycloalkyl or 4-10    membered heterocycloalkyl ring, each optionally substituted with 1,    2, 3, 4, or 5 substituents independently selected from Cy², Cy²-C₁₋₄    alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl,    CN, NO₂, OR^(a1), SR^(a1), C(O)R^(b1), C(O)NR^(c1)R^(d1),    C(O)OR^(a1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1), NR^(c1)R^(d1),    NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1), NR^(c1)C(O)NR^(c1)R^(d1),    C(═NR^(e1))R^(b1), C(═NR^(e1))NR^(c1)R^(d1),    NR^(c1)C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)S(O)R^(b1),    NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1),    S(O)NR^(c1)R^(d1), S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);

-   R⁴ is H, C₁₋₄ alkyl, halo, C₁₋₄ haloalkyl, or CN;

-   R⁵ is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₁₀    cycloalkyl, 5-14 membered heteroaryl, 4-14 membered    heterocycloalkyl, C₆₋₁₀ aryl —C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄    alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered    heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a2), SR^(a2), C(O)R^(b2),    C(O)NR^(c2)R^(d2), C(O)OR^(a2), OC(O)R^(b2), OC(O)NR^(c2)R^(d2),    NR^(c2)R^(d2) NR^(c2)C(O)R^(b2), NR^(c2)C(O)OR^(a2),    NR^(c2)C(O)NR^(c2)R^(d2), C(═NR^(e2))R^(b2),    C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)C(═NR^(e2))NR^(c2)R^(d2),    NR^(c2)S(O)R^(b2), NR^(c2)S(O)₂R^(b2), NR^(c2)S(O)₂NR^(c2)R^(d2),    S(O)R^(b2), S(O)NR^(c2)R^(d2), S(O)₂R^(b2), and S(O)₂NR^(c2)R^(d2);    wherein said C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl,    C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl, 5-14 membered heteroaryl, 4-14    membered heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇    cycloalkyl-C₁₋₄ alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and 4-10    membered heterocycloalkyl-C₁₋₄ alkyl of R¹ are each optionally    substituted with 1, 2, 3, 4, or 5 substituents independently    selected from Cy³, Cy³-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl,    C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a2), SR^(a2), C(O)R^(b2),    C(O)NR^(c2)R^(d2), C(O)OR^(a2), OC(O)R^(b2), OC(O)NR^(c2)R^(d2),    NR^(c2)R^(d2), NR^(c2)C(O)R^(b2), NR^(c2)C(O)OR^(a2),    NR^(c2)C(O)NR^(c2)R^(d2), C(═NR^(e2))R^(b2),    C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)C(═NR^(e2))NR^(c2)R^(d2),    NR^(c2)S(O)R^(b2), NR^(c2)S(O)₂R^(b2), NR^(c2)S(O)₂NR^(c2)R^(d2),    S(O)R^(b2), S(O)NR^(c2)R^(d2), S(O)₂R^(b2), or S(O)₂NR^(c2)R^(d2);

-   each R⁶ is independently selected from H, halo, C₁₋₆ alkyl, C₁₋₆    haloalkyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl, 4-7    membered heterocycloalkyl, CN, NO₂, OR^(a3), SR^(a3), C(O)R^(b3),    C(O)NR^(c3)R^(d3), C(O)OR^(a3), OC(O)R^(b3), OC(O)NR^(c3)R^(d3),    NR^(c3)R^(d3), NR^(c3)C(O)R^(b3), NR^(c3)C(O)NR^(c3)R^(d3),    NR^(c3)C(O)OR^(a3), C(═NR^(e3))NR^(c3)R^(d3),    NR^(c3)C(═NR^(e3))NR^(c3)R^(d3), S(O)R^(b3), S(O)NR^(c3)R^(d3),    S(O)₂R^(b3), NR^(c3)S(O)₂R^(b3), NR^(c3)S(O)₂NR^(c3)R^(d3), and    S(O)₂NR^(c3)R^(d3), wherein said C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₆₋₁₀    aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl, 4-7 membered    heterocycloalkyl of R⁶ are each optionally substituted with 1, 2, 3,    4, or 5 substituents independently selected from halo, C₁₋₆ alkyl,    C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a3),    SR^(a3), C(O)R^(b3), C(O)NR^(c3)R^(d3), C(O)OR^(a3), OC(O)R^(b3),    OC(O)NR^(c3)R^(d3), NR^(c3)R^(d3), NR^(c3)C(O)R^(b3),    NR^(c3)C(O)NR^(c3)R^(d3), NR^(c3)C(O)OR^(a3),    C(═NR^(e3))NR^(c3)R^(d3), NR^(c3)C(═NR^(e3))NR^(c3)R^(d3),    S(O)R^(b3), S(O)NR^(c3)R^(d3), S(O)₂R^(b3), NR^(c3)S(O)₂R^(b3),    NR^(c3)S(O)₂NR^(c3)R^(d3), and S(O)₂NR^(c3)R^(d3);

-   each Cy¹ is independently selected from C₆₋₁₀ aryl, C₃₋₁₀    cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered    heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or 5    substituents independently selected from halo, C₁₋₆ alkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl,    5-14 membered heteroaryl, 4-14 membered heterocycloalkyl, C₆₋₁₀    aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered    heteroaryl-C₁₋₄ alkyl, 4-10 membered heterocycloalkyl-C₁₋₄ alkyl,    CN, NO₂, OR^(a), SR^(a), C(O)R^(b), C(O)NR^(c)R^(d), C(O)OR^(a),    OC(O)R^(b), OC(O)NR^(c)R^(d), NR^(c)R^(d), NR^(c)C(O)R^(b),    NR^(c)C(O)OR^(a), NR^(c)C(O)NR^(c)R^(d), C(═NR^(e))R^(b),    C(═NR^(e))NR^(c)R^(d), NR^(c)C(═NR^(e))NR^(c)R^(d), NR^(c)S(O)R^(b),    NR^(c)S(O)₂R^(b), NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b),    S(O)NR^(c)R^(d), S(O)₂R^(b), and S(O)₂NR^(c)R^(d);

-   each Cy² is independently selected from C₆₋₁₀ aryl, C₃₋₁₀    cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered    heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or 5    substituents independently selected from halo, C₁₋₆ alkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl,    5-14 membered heteroaryl, 4-14 membered heterocycloalkyl, C₆₋₁₀    aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered    heteroaryl-C₁₋₄ alkyl, 4-10 membered heterocycloalkyl-C₁₋₄ alkyl,    CN, NO₂, OR^(a1), SR^(a1), C(O)R^(b1), C(O)NR^(c1)R^(d1),    C(O)OR^(a1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1), NR^(c1)R^(d1),    NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1), NR^(c1)C(O)NR^(c1)R^(d1),    C(═NR^(e1))R^(b1), C(═NR^(e1))NR^(c1)R^(d1),    NR^(c1)C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)S(O)R^(b1),    NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1),    S(O)NR^(c1)R^(d1), S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);

-   each Cy³ is independently selected from C₆₋₁₀ aryl, C₃₋₁₀    cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered    heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or 5    substituents independently selected from halo, C₁₋₆ alkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl,    5-14 membered heteroaryl, 4-14 membered heterocycloalkyl, C₆₋₁₀    aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered    heteroaryl-C₁₋₄ alkyl, 4-10 membered heterocycloalkyl-C₁₋₄ alkyl,    CN, NO₂, OR^(a2), SR^(a2), C(O)R^(b2), C(O)NR^(c2)R^(d2),    C(O)OR^(a2), OC(O)R^(b2), OC(O)NR^(c2)R^(d2), NR^(c2)R^(d2),    NR^(c2)C(O)R^(b2), NR^(c2)C(O)OR^(a2), NR^(c2)C(O)NR^(c2)R^(d2),    C(═NR^(e2))R^(b2), C(═NR^(e2))NR^(c2)R^(d2),    NR^(c2)C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)S(O)R^(b2),    NR^(c2)S(O)₂R^(b2), NR^(c2)S(O)₂NR^(c2)R^(d2), S(O)R^(b2),    S(O)NR^(c2)R^(d2), S(O)₂R^(b2), and S(O)₂NR^(c2)R^(d2);

-   each R^(a), R^(b), R^(c), R^(d), R^(a1), R^(b1), R^(c1), R^(d1),    R^(a2), R^(b2), R^(c2), R^(d2), R^(a3), R^(b3), R^(c3), and R^(d3)    is independently selected from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-10 membered    heteroaryl, 4-10 membered heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl,    C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and    4-10 membered heterocycloalkyl-C₁₋₄ alkyl, wherein said C₁₋₆ alkyl,    C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-10    membered heteroaryl, 4-10 membered heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄    alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered heteroaryl-C₁₋₄    alkyl, and 4-10 membered heterocycloalkyl-C₁₋₄ alkyl of R^(a),    R^(b), R^(c), R^(d), R^(a1), R^(b1), R^(c1), R^(d1), R^(a2), R^(b2),    R^(c2), R^(d2), R^(a)R^(b3), R^(c3), or R^(d3) is optionally    substituted with 1, 2, 3, 4, or 5 substituents independently    selected from halo, C1-4 alkyl, C1-4 haloalkyl, Ci-₆ haloalkyl, C2-6    alkenyl, C2-6 alkynyl, CN, OR^(a4), SR^(a4), C(O)R^(b4),    C(O)NR^(c4)R^(d4), C(O)OR^(a4), OC(O)R^(b4), OC(O)NR^(c4)R^(d4),    NR^(c4)R^(d4), NR^(c4)C(O)R^(b4), NR^(c4)C(O)NR^(c4)R^(d4),    NR^(c4)C(O)OR^(a4), C(═NR^(e4))NR^(c4)R^(d4),    NR^(c4)C(═NR^(e4))NR^(c4)R^(d4), S(O)R^(b4), S(O)NR^(c4)R^(d4),    S(O)₂R^(b4), NR^(c4) S(O)₂R^(m), NR^(c4)S(O)₂NR^(c4)R^(d4), and    S(O)₂NR^(c4)R^(d4);

-   each R^(a4), R^(b4), R^(c4), and R^(d4) are independently selected    from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,    C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl, 4-7 membered    heterocycloalkyl, wherein said C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6 membered    heteroaryl, and 4-7 membered heterocycloalkyl are each optionally    substituted with 1, 2, or 3 substituents independently selected from    OH, CN, amino, halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, and    C₁₋₆ haloalkoxy;    -   each R^(e), R^(e1), R^(e2), R^(e3), and R^(e4) is independently        selected from H, C₁₋₄ alkyl, and CN;

-   R_(PTM) is H; halogen (e.g., Cl or F); —CN; —OH; —NO₂; —NH₂;    optionally substituted linear or branched alkyl (e.g., optionally    substituted linear or branched C1-C6 alkyl or optionally substituted    linear or branched C1-C4 alkyl or C1-C8 alkyl optionally substituted    with OH); optionally substituted cycloalkyl (e.g., cyclopropyl,    cyclobutyl, cyclopentyl, cyclohexyl); O-optionally substituted    linear or branched C1-C4 alkyl; an optionally substituted C1-C4    alkynyl; an optionally substituted C1-C4 alkyne; optionally    substituted linear or branched hydroxyalkyl (e.g., optionally    substituted linear or branched C1-C7 hydroxy alkyl); optionally    substituted alkylcycloalkyl (e.g., includes optionally substituted    C1-C6 alkyl, optionally substituted C3-C10 cycloalkyl; or both);    optionally substituted alkyl-aryl (e.g., includes an optionally    substituted linear or branched C1-C6 alkyl, an optionally    substituted 5-10 member heteroaryl, or both); optionally substituted    alkyl-heteroaryl (e.g., includes an optionally substituted linear or    branched C1-C6 alkyl, an optionally substituted 5-10 member    heteroaryl, or both); optionally substituted alkyl-heteroaryl (e.g.,    includes a C1-C6 alkyl, an optionally substituted 5 or 6 member    heteroaryl, optionally substituted with a C1-C4 alkyl; the    heteroaryl is selected from oxazol-4-yl, 1,3,4-triazol-2-yl, and    imidazole-1-yl; or combination thereof); optionally substituted    —NH-alkyl-heteroaryl (e.g., an optionally substituted linear or    branched C1-C5 alkyl, an optionally substituted 5-8 member    heteroaryl, optionally substituted with a C1-C4 alkyl,    N—CH₂-pyrazol-4-yl, or a combination thereof); optionally    substituted alkoxy (e.g., an optionally substituted linear or    branched C1-C6 alkyl or —OCH₃); optionally substituted    O-heterocyclyl (e.g., includes an optionally substituted 3-12 or 4-7    member heterocyclyl; an optionally substituted heterocycloalkyl; an    optionally substituted C₃₋₁₂ monocyclic or bicyclic    heterocycloalkyl; optionally substituted with at least one OH, C1-C5    alkyl (such as a methyl), ═O, NH₂, or a combination thereof; or a    combination thereof); optionally substituted S-heterocyclyl (e.g.,    includes an optionally substituted 4-7 member heterocyclyl; an    optionally substituted heterocycloalkyl; optionally substituted with    at least one C1-C4 alkyl (such as a methyl), ═O, or a combination    thereof; or a combination thereof); optionally substituted

(e.g., optionally substituted with a linear or branched C1-C4 alkyl;—(CH₂)_(u)CO(CH₂)_(v)CH₃, —COCH₃, or —CH₂CH₂COCH₃, wherein each u and vis independently selected from 1, 2, 3, 4 or 5);

-   -   optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —O(CH₂)_(u)CO(CH₂)_(v)CH₃,        —O(CH₂)_(u)CH((CH₂)_(x)CH₃)(CH₂)_(w)CO(CH₂)_(v)CH₃, —O—CH₂COCH₃,        —O—CH₂COCH₂CH₃, —O—CH(CH₃₎COCH₃, —OCH₂COCH₃, or —OCH₂(CH₃)COCH₃,        wherein each u, v, w, and * is independently selected from 1, 2,        3, 4 or 5); optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a),        —CONR_(PTM1a)R_(PTM2a), —CH₂CONR_(PTM1a)R_(PTM2a),        —CH₂CH₂CONR_(PTM1a)R_(PTM2a), —CONHCH₃, or —CH₂CONHCH₃, wherein        each u and v is independently selected from 1, 2, 3, 4 or 5);        optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —O(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a),        —O(CH₂)_(u)CH((CH₂)_(x)CH₃)(CH₂)_(w)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a),        —O—CH(CH₃)CONR_(PTM1a)R_(PTM2a), —O—CH₂CONR_(PTM1a)R_(PTM2a), or        —OCH₂C(O)NHOCH₃, wherein each u, v, w, and x is independently        selected from 1, 2, 3, 4 or 5); optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —(CH₂)_(u)CHCH(CH₂)_(w)CO(CH₂)_(v) NR_(PTM1a)R_(PTM2a) or        —CHCHCONR_(PTM1a)R_(PTM2a), wherein each u, v, and w is        independently selected from 1, 2, 3, 4 or 5); optionally        substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —NH—(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a) or        —NH—CH₂CONR_(PTM1a)R_(PTM2a), wherein each u and v is        independently selected from 1, 2, 3, 4 or 5); fluoroalkoxy        (e.g., a mono-, bi- and/or tri-fluoroalkoxy); optionally        substituted monocyclic or bicyclic cycloalkyl (e.g., an        optionally substituted 3-12 member cycloalkyl; optionally        substituted with at least one of OH, ═O, linear or branched        C1-C6 alkyl (such as a methyl, ethyl, or butyl), or NH₂; or a        combination thereof); optionally substituted hydroxycycloalkyl;        optionally substituted aryl (e.g., an optionally substitute        C5-C10 aryl, an optionally substituted 5-7 member aryl;        optionally substituted with at least one halogen or C1-C3 alkyl        (e.g., methyl or ethyl); or a combination thereof), optionally        substituted heteroaryl (e.g., an optionally substituted 5-10 or        member heteroaryl, an optionally substituted 5-7 member        heteroaryl; an optionally substituted 5-member heteroaryl;        optionally substituted with at least one halogen or C1-C3 alkyl        (e.g., methyl or ethyl); or a combination thereof) optionally        linked to Q via a C or N-atom of the heteroaryl (e.g., at least        one of optionally linked to Q, optionally linked via an        optionally substituted —(CH₂)_(u)O(CH₂)_(v)O(CH₂)_(x)—, or a        combination thereof); optionally substituted monocyclic or        bicyclic heterocyclyl (e.g., an optionally substituted 3-12        member heterocyclyl; an C3-C12 monocyclic or bicyclic        heterocycloalkyl, azetidine1-yl, pyrrolidin-1-yl, piperidin-1yl,        piperazin-1-yl, or morpholin-4-yl, or homopiperazin-1-yl, each        optionally substituted with OH, a linear or branched C1-C5 alkyl        (a methyl, ethyl, or butyl group) or NH₂) optionally linked to Q        via a C or N atom of the heterocyclyl (e.g., at least one of        optionally linked to Q, optionally linked via an optionally        substituted —(CH₂)_(u)O(CH₂)_(v)O(CH₂)_(x)—, or both);

-   t₁ is selected from 1, 2, 3, 4, or 5;

-   each t₂ is independently is independently selected from 0, 1, 2, 3,    4, or 5;

-   R_(PTM1a) and R_(PTM2a) are independently H, optionally substituted    C1-C4 alkyl (e.g., a CH₃ or CH₂CH₃), optionally substituted C1-C4    alkoxy (e.g., —OCH₂ or —CH₂CH₃), optionally substituted CH₂OCH₃ or    R_(PTM1a) and R_(PTM2a) are joined together form an optionally    substituted 3-10 member ring;

-   n is an integer from 0 to 10; and

-   of the PTM indicates the point of attachment with a chemical linker    group or a ULM.

In any of the aspects or embodiments described herein, R_(PTM) of PTM-Iis modified to be covalently linked to a linker group (L) or a ULM (suchas CLM). In any aspect or embodiment described herein, R_(PTM) of PTM-Iis modified to be covalently linked to a chemical linker group (L) or aULM.

In any aspect or embodiment described herein, W of PTM-I is O.

In any aspect or embodiment described herein, Q of PTM-I is group (a)and ring B is selected from:

wherein

-   R¹, R^(1A), R^(1B), R^(1C) and R^(1D) are each independently    selected from H, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆    haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl, 5-14 membered heteroaryl,    4-14 membered heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇    cycloalkyl-C₁₋₄ alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10    membered heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, and OR^(a), SR^(a),    C(O)R^(b), C(O)NR^(c)R^(d), C(O)OR^(a), OC(O)R^(b),    OC(O)NR^(c)R^(d), NR^(c)R^(d), NR^(c)C(O)R^(b), NR^(c)C(O)OR^(a),    NR^(c)C(O)NR^(c)R^(d), C(═NR^(e))R^(b), C(═NR^(e))NR^(c)R^(d),    NR^(c)C(═NR^(e))NR^(c)R^(d), NR^(c)S(O)R^(b), NR^(c)S(O)₂R^(b),    NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b), S(O)NR^(c)R^(d), S(O)₂R^(b),    S(O)₂NR^(c)R^(d), and R_(PTM), wherein said C₁₋₆ alkyl, C₂₋₆    alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀ cycloalkyl,    5-14 membered heteroaryl, 4-14 membered heterocycloalkyl, C₆₋₁₀    aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered    heteroaryl-C₁₋₄ alkyl, 4-10 membered heterocycloalkyl-C₁₋₄ alkyl,    wherein one of R¹, R^(1A), R^(1B), R^(1C), and R^(1D) is optionally    substituted with 1, 2, 3, 4, or 5 substituents independently    selected from Cy¹, Cy¹-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl,    C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a), SR^(a), C(O)R^(b),    C(O)NR^(c)R^(d), C(O)OR^(a), OC(O)R^(b), OC(O)NR^(c)R^(d),    NR^(c)R^(d), NR^(c)C(O)R^(b), NR^(c)C(O)OR^(a),    NR^(c)C(O)NR^(c)R^(d), C(═NR^(e))R^(b), C(═NR^(e))NR^(c)R^(d),    NR^(c)C(═NR^(e))NR^(c)R^(d), NR^(c)S(O)R^(b), NR^(c)S(O)₂R^(b),    NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b), S(O)NR^(c)R^(d), S(O)₂R^(b), and    S(O)₂NR^(c)R^(d);-   or R^(1A) and R^(1B) together form a C₃₋₇ cycloalkyl or 4-10    membered heterocycloalkyl ring, each optionally substituted with 1,    2, 3, 4, or 5 substituents independently selected from Cy¹, Cy¹C₁₋₄    alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl,    CN, NO₂, OR^(a), SR^(a), C(O)R^(b), C(O)NR^(c)R^(d), C(O)OR^(a),    OC(O)R^(b), OC(O)NR^(c)R^(d), NR^(c)R^(d), NR^(c)C(O)R^(b),    NR^(c)C(O)OR^(a), NR^(c)C(O)NR^(c)R^(d), C(═NR^(e))R^(b),    C(═NR^(e))NR^(c)R^(d), NR^(c)C(═NR^(e))NR^(c)R^(d), NR^(c)S(O)R^(b),    NR^(c)S(O)₂R^(b), NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b),    S(O)NR^(c)R^(d), S(O)₂R^(b), S(O)₂NR^(c)R^(d), and R_(PTM), wherein    ring B or a substituent of ring B is covalently coupled to at least    one of R_(PTM), a linking group (L) or a CLM.

In any aspect or embodiment described herein, Q if PTM-I is group (a)and ring B is selected from:

wherein R_(PTM) is as described herein, and the

indicates a covalent attachment to a linking group (L) or CLM.

In any of the aspects or embodiments, the Q of PTM-I is covalentlycoupled to the linker (L) or CLM.

In any aspect or embodiment described herein, Q of PTM-I is group (a):

wherein A¹, A², and A³ are CR⁶; R⁶ is independently selected from H,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6membered heteroaryl, 4-7 membered heterocycloalkyl, CN, and NO₂; ring Bis a group as described herein, the

of Q indicates the point of attachment with the nitrogen, and the

of Q indicates the site of attachment to at least one of R_(PTM), achemical linking group (L) or CLM. In any of the aspects or embodiments,the Q of PTM-I is covalently coupled to a chemical linking group (L) orCLM.

In any aspect or embodiment described herein, Q of PTM-I is:

wherein A¹, A² and A³ are defined in any aspect or embodiment describedherein, the

of Q indicates the point of attachment with the nitrogen, and the

indicates the site of covalent attachment to at least one of R_(PTM), achemical linking group (L) or CLM. In any of the aspects or embodiments,the Q of PTM-I is covalently coupled to the linker (L) or CLM.

In any aspect or embodiment described herein, Q of PTM-I is:

wherein A¹, A², and A³ are defined in any aspect or embodiment describedherein, the

of Q indicates the point of attachment with the nitrogen, and the

indicates the site of covalent attachment to at least one of R_(PTM), achemical linking group (L) or CLM. In any of the aspects or embodiments,the Q of PTM-I is covalently coupled to the linker (L) or CLM.

In any aspect or embodiment described herein, Q of PTM-I is:

wherein:

-   -   A¹ and A² are each CR⁶;    -   each R⁶ is independently selected from H, halo, C₁₋₆ alkyl, C₁₋₆        haloalkyl, Ono aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl,        4-7 membered heterocycloalkyl, CN, and NO₂;    -   the        of Q indicates the point of attachment with the nitrogen; and    -   the        indicates a covalent attachment to at least one of R_(PTM), a        chemical linking group (L) or CLM.

In any aspect or embodiment described herein, W PTM-I is O.

In any aspect or embodiment described herein, R² PTM-I is H.

In any aspect or embodiment described herein, R⁵ of PTM-I is H or C₁₋₆alkyl.

In any aspect or embodiment described herein, R^(3A) and R^(3B) of PTM-Iare each independently selected from H, halo, and C₁₋₆ alkyl.

In any aspect or embodiment described herein, R_(PTM) is selected from abond, methyl, ethyl, propyl, butyl, pentyl, hexyl, H, O H, ethyl,

wherein

represents the site of attachment with PTM and the or

indicates the site of covalent attachment with a chemical linking group(L) or CLM.

In any of the aspects or embodiments, the R_(PTM) is covalently coupledto the linker (L) or CLM.

In any aspect or embodiment described herein, the PTM-I is selected fromthe group consisting of:

wherein R_(PTM) is a bond, an atom or chemical group as describedherein, and the

of the R_(PTM) indicates the point of attachment of a chemical linkinggroup (L) or a CLM.

In any aspect or embodiment described herein, R_(PTM) is a 5-7 memberedaryl or heteroaryl (e.g., a 6-membered aryl or heteroaryl), wherein:

-   -   the heteroaryl has 1, 2, or 3 heteroatoms selected from O, N,        and S; and    -   the R_(PTM) is optionally substituted with (a) Cy¹ or (b) a 3-        to 10-membered cycloalkyl (e.g., 5- or 6-membered cycloalkyl),        heterocycloalkyl (e.g., 5- or 6-membered heterocycloalkyl),        spirocycloalky (e.g., 6- to 12-membered or 8- to 10-membered        spirocycloalkyl), spiroheterocycloalkyl (e.g., 6- to 12-membered        or 8- to 10-membered spiroheterocycloalkyl), bicyclic cycloalkyl        (e.g., 6- to 10-membered bicyclic cycloalkyl), or bicyclic        hetercycloalkyl (5- to 10-membered bicyclic heterocycloalkyl),        wherein each of these is optionally substituted with 1, 2, 3, or        4 groups selected from the group consisting of H, C₁₋₃ alkyl,        methyl, ethyl, and halogen.

In any aspect or embodiment described herein, R_(PTM) is a 5-7 memberedaryl or heteroaryl (e.g., a 6-membered aryl or heteroaryl), wherein:

-   -   the heteroaryl has 1, 2, or 3 heteroatoms selected from O, N,        and S; and    -   the R_(PTM) is optionally substituted with a 3- to 10-membered        cycloalkyl (e.g., 5- or 6-membered cycloalkyl), heterocycloalkyl        (e.g., 5- or 6-membered heterocycloalkyl), spirocycloalky (e.g.,        6- to 12-membered spirocycloalkyl), spiroheterocycloalkyl (e.g.,        6- to 12-membered spiroheterocycloalkyl), bicyclic cycloalkyl        (e.g., 6- to 10-membered bicyclic cycloalkyl), or bicyclic        hetercycloalkyl (5- to 10-membered bicyclic heterocycloalkyl),        wherein each of these is optionally substituted with 1, 2, 3, or        4 groups selected from the group consisting of H, C₁₋₃ alkyl,        methyl, ethyl, and halogen.

In any aspect or embodiment described herein R_(PTM1a) or R_(PTM2a) isselected from: a bond, methyl, ethyl, propyl, butyl, pentyl, hexyl, H, OH, ethyl,

wherein

represents the site of attachment with PTM and the

or * indicates the site of covalent attachment with a chemical linkinggroup (L) or CLM.

In any aspect or embodiment described herein, the R_(PTM) or thecorresponding location of any PTM described herein (e.g. PTM-I andderivatives thereof) is a linear or branched C1-C8 alkyl optionallysubstituted with OH.

In any aspect or embodiment described herein, the R_(PTM) or thecorresponding location of any PTM described herein (e.g. PTM-Iderivatives thereof) is H, OH, CN, optionally substituted linear orbranched C1-C4 alkyl, O-optionally substituted linear or branched C1-C4alkyl, an optionally substituted C1-C4 alkynyl, an optionallysubstituted C1-C4 alkyne, an optionally substituted monocyclic orbicyclic C3-C12 heterocyclyl (e.g., an optionally substituted C3-C12monocyclic or bicyclic heterocycloalkyl, such as an C3-C12 monocyclic orbicyclic heterocycloalkyl, azetidine1-yl, pyrrolidin-1-yl,piperidin-1yl, piperazin-1-yl, or morpholin-4-yl, or homopiperazin-1-yl,each optionally substituted with one or more of OH, a linear or branchedC1-C5 alkyl or NH₂), or an optionally substituted —O—C₃₋₁₂ monocyclic orbicyclic heterocyclyl (e.g., an optionally substituted —O—C₃₋₁₂monocyclic or bicyclic heterocycloalkyl, such as —O—C₃₋₁₂ monocyclic orbicyclic heterocycloalkyl optionally substituted with at least one OH, alinear or branched C1-C5 alkyl or NH₂), or an optionally substitutedC3-C12 member ring (e.g., an optionally substituted C3-C12 non-arylmembered ring optionally substituted with one or more of OH, linear orbranched C1-C5 alkyl, or NH₂), wherein when R_(PTM) is a ring structureit is optionally covalently linked to Q₁₆ via a C or N of the R_(PTM)ring.

In any aspect or embodiment described herein, the PTM is represented bythe chemical structure:

wherein X, A¹, A², A³, R², R^(3A), R^(3B), R⁴, R⁵, and R_(PTM) aredefined as described in any aspect or embodiment herein, and the

indicates a covalent attachment to a chemical linking group (L) or CLM

In any aspect or embodiment described herein, the PTM is represented bythe chemical structure:

wherein R², R^(3A), R^(3B), R⁴, R⁵, and R_(PTM) are defined as in anyaspect or embodiment described herein, and the

indicates a covalent attachment to a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is represented by a chemical structure selectedfrom:

wherein the

indicates a covalent attachment of a chemical linking group (L) or CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) is selected from:

wherein the

of the PTM indicates the site of attachment of a chemical linking group(L) or a CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) has the chemical structure:

and wherein the

of the PTM indicates the covalent attachment of a chemical linking groupor a CLM.

In any aspect or embodiment described herein, the PTM (for e.g. PTM-I,PTM-IIa, PTM-IIb, PTM-IIIa, or PTM-IIIb, or the PTM of FormulasIIIa-IIIh and IVa-IVh) has the chemical structure:

wherein the

indicates a site of attachment of a linker or ULM, and wherein each PTMis coupled to at least one linker or ULM

In any aspect or embodiment described herein, the hetero-bifunctionalcompound is represented by the chemical structure:

wherein:

R^(3A), R^(3B), R⁴ and R⁵ are defined as described herein;

-   -   X is CH or N;    -   R⁷ is H, C1-C6 alkyl, C1-C6 alkoxy, or halogen;    -   A is a 4-7 membered aryl, heteroaryl, cycloalkyl or        heterocycloalkyl;    -   L is a chemical linker group optionally coupled to the        phthalimido group via an oxygen, amine group or methyl group;    -   X is CH or N; and    -   Y is O or H₂ (i.e., absent).

In any aspect or embodiment described herein, the hetero-bifunctionalcompound is represented by the chemical structure:

wherein:

-   -   Z₁ is an R group of a CLM as described in any aspect or        embodiment described herein that is modified to be covalently        linked to L, such a group selected from —C(═O)—, —CONR′—, —O—,        —NR′—, a carbon shared with a cyclic group of L, or a nitrogen        shared with a cyclic group of L;    -   n is an integer from 0 to 3 (e.g., 0, 1, 2, or 3);    -   R is selected from H, O, OH, N, NH, NH₂, Cl, —F, —Br, —I,        methyl, optionally substituted linear or branched alkyl (e.g.,        optionally substituted linear or branched C1-C6 alkyl),        optionally substituted linear or branched alkoxy (e.g.,        optionally substituted linear or branched C1-C6 alkoxy),        -alkyl-aryl (e.g., an -alkyl-aryl comprising at least one of        C1-C6 alkyl, C4-C7 aryl, or a combination thereof), aryl (e.g.,        C5-C7 aryl), amine, amide, or carboxy;    -   L, R^(3A), R^(3B), R⁴, and R⁵ are defined as in any aspect or        embodiment described herein;    -   each X is individually CH or N;    -   R⁷ is H, C1-C6 alkyl, C1-C6 alkoxy, or halogen;    -   A is a 4-7 membered aryl, heteroaryl, cycloalkyl or        heterocycloalkyl or a 6- to 12-membered (e.g., 8- to        10-membered) spirocycloalkyl or spiroheterocycloalkyl, each        optionally substituted with 1, 2, 3, or 4 groups selected from        the group consisting of H, C₁₋₃ alkyl, methyl, ethyl, and        halogen;    -   X is CH or N;    -   A¹, A², and A³ as defined in any aspect or embodiment described        herein (e.g., A¹ and A³ are individually N or CH, and A² is CR⁷        or N); and    -   Y is O or H₂ (i.e., absent).

In any aspect or embodiment described herein, the hetero-bifunctionalcompound is represented by the chemical structure:

wherein:

-   -   R^(3A), R^(3B), R⁴, and R⁵ are defined as described herein;    -   R⁷ is H or halogen;    -   A is a 4-7 membered heterocycloalkyl;    -   L is a chemical linker group optionally coupled to the        phthalimido group via an oxygen, amine group or methyl group;    -   X is CH or N; and    -   Y is O or H₂ (i.e., absent).

In any aspect or embodiment described herein, the hetero-bifunctionalcompound is represented by the chemical structure:

wherein:

-   -   Z₁ is an R group of a CLM as described in any aspect or        embodiment described herein that is modified to be covalently        linked to L, such a group selected from —C(═O)—, —CONR′—, —O—,        —NR′—, a carbon shared with a cyclic group of L, or a nitrogen        shared with a cyclic group of L;    -   n is an integer from 0 to 3 (e.g., 0, 1, 2, or 3);    -   R is selected from H, O, OH, N, NH, NH₂, Cl, —F, —Br, —I,        methyl, optionally substituted linear or branched alkyl (e.g.,        optionally substituted linear or branched C1-C6 alkyl),        optionally substituted linear or branched alkoxy (e.g.,        optionally substituted linear or branched C1-C6 alkoxy),        -alkyl-aryl (e.g., an -alkyl-aryl comprising at least one of        C1-C6 alkyl, C4-C7 aryl, or a combination thereof), aryl (e.g.,        C5-C7 aryl), amine, amide, or carboxy;    -   L, R^(3A), R^(3B), R⁴, and R⁵ are defined as in any aspect or        embodiment described herein;    -   R⁷ is H or halogen;    -   A is a 4-7 membered heterocycloalkyl or a 6- to 12-membered        (e.g., 8- to 10-membered) spiroheterocycloalkyl, each optionally        substituted with 1, 2, 3, or 4 groups selected from the group        consisting of H, C₁₋₃ alkyl, methyl, ethyl, and halogen;    -   each X is individually CH or N;    -   A¹, A², and A³ as defined in any aspect or embodiment described        herein (e.g., A¹ and A³ are individually N or CH, and A² is CR⁷        or N); and    -   Y is O or H₂ (i.e., absent).

In any aspect or embodiment described herein, the hetero-bifunctionalcompound has the chemical structure:PTM-L-CLM,

or a pharmaceutically acceptable salt or solvate thereof,

wherein:

(a) the CLM is a small molecule E3 ubiquitin ligase binding moiety thatbinds a cereblon E3 ubiquitin ligase and is represented by the chemicalstructure:

wherein:

-   -   W is selected from the group consisting of CH₂, O, CHR, C═O,        SO₂, NH, N, optionally substituted cyclopropyl group, optionally        substituted cyclobutyl group, and N-alkyl;    -   Q₁, Q₂, Q₃, Q₄, Q₅ each independently represent a carbon C or N        substituted with a group independently selected from R′, N or        N-oxide;    -   R¹ is selected from absent (i.e., a bond), H, OH, CN, C1-C3        alkyl, C═O;    -   R² is selected from the group absent (i.e., a bond), H, OH, CN,        C1-C3 alkyl, CHF₂, CF₃, CHO, C(═O)NH₂;    -   R³ is selected from a bond, H, alkyl (e.g., C1-C6 or C1-C3        alkyl), substituted alkyl (e.g., substituted C1-C6 or C1-C3        alkyl), alkoxy (e.g., C1-C6 or C1-C3 alkoxyl), substituted        alkoxy (e.g., substituted C1-C6 or C1-C3 alkoxyl);    -   R⁴ is selected from a bond, H, alkyl, substituted alkyl;    -   R⁵ and R⁶ are each independently a bond, H, halogen, C(═O)R′,        CN, OH, CF₃;    -   X is C, CH, C═O, or N;    -   X₁ is C═O, N, CH, or CH₂;    -   R′ is selected from a bond, H, halogen, amine, alkyl (e.g.,        C1-C3 alkyl), substituted alkyl (e.g., substituted C1-C3 alkyl),        alkoxy (e.g., C1-C3 alkoxyl), substituted alkoxy (e.g.,        substituted C1-C3 alkoxyl), NR²R³, C(═O)OR², optionally        substituted phenyl;    -   n is 0-4;    -   is a single or double bond; and    -   the CLM is covalently joined to a PTM, a chemical linker group        (L), a ULM, or CLM);        (b) the PTM is a small molecule Leucine-rich repeat kinase 2        (LRRK2) targeting moiety that binds to the human LRRK2 or mutant        thereof represented by the chemical structure:

wherein:

-   -   X is CH or N;    -   W is O or S;    -   Q is selected from one of the following:

-   -    wherein:        -   A¹, A², and A³ are each independently selected from N and            CR⁶, wherein for (a) no more than two of A¹, A², and A³ are            simultaneously N,        -   ring B is an optionally substituted C3-C8 aryl, optionally            substituted C3-C8 heteroaryl, optionally substituted C3-C8            cycloalkyl, optionally substituted C3-C8 heterocycloalkyl,            optionally substituted C3-C8 aryl, optionally substituted            C3-C8 aryl,        -   the            of Q indicates the point of attachment with the nitrogen,            and        -   the            of Q indicates an optional covalent attachment to R_(PTM);        -   R² is H or C₁₋₄ alkyl;    -   R^(3A) and R^(3B) are each independently selected from H, halo,        C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀        aryl, C₃₋₁₀ cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a1), SR^(a1),        C(O)R^(b1), C(O)NR^(c1)R^(d1), C(O)OR^(a1), OC(O)R^(b1),        OC(O)NR^(c1)R^(d1), NR^(d)R^(d1), NR^(c1)C(O)R^(b1),        NR^(c1)C(O)OR^(a1), NR^(c1)C(O)NR^(c1)R^(d1), C(═NR^(e1))R^(b1),        C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)(═NR^(e1))NR^(c1)R^(d1),        NR^(c1)S(O)R^(b1), NR^(c1)S(O)₂R^(b1),        NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1), S(O)NR^(c1)R^(d1),        S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1); wherein said C₁₋₆ alkyl,        C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl of R¹ are each optionally        substituted with 1, 2, 3, 4, or 5 substituents independently        selected from Cy², Cy²-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆        alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a1),        SR^(a1), C(O)R^(b1), C(O)NR^(c1)R^(d1), C(O)OR^(a1),        OC(O)R^(b1), OC(O)NR^(c1)R^(d1), NR^(c1)R^(d1),        NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1), NR^(c1)C(O)NR^(c1)R^(d1),        C(═NR^(e1))R^(b1), C(═NR^(e1))NR^(c1)R^(d1),        NR^(c1)C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)S(O)R^(b1),        NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1),        S(O)NR^(c1)R^(d1), S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);    -   or R^(3A) and R^(3B) together form a C₃₋₇ cycloalkyl or 4-10        membered heterocycloalkyl ring, each optionally substituted with        1, 2, 3, 4, or 5 substituents independently selected from Cy²,        Cy²-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,        C₁₋₆ haloalkyl, CN, NO₂, OR^(a1), SR^(a1), C(O)R^(b1),        C(O)NR^(c1)R^(d1), C(O)OR^(a1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1),        NR^(c1)R^(d1), NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1),        NR^(c1)C(O)NR^(c1)R^(d1), C(═NR^(e1))R^(b1),        C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)C(═NR^(e1))NR^(c1)R^(d1),        NR^(c1)S(O)R^(b1), NR^(c1)S(O)₂R^(b1),        NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1), S(O)NR^(c1)R^(d1),        S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);    -   R⁴ is H, C₁₋₄ alkyl, halo, C₁₋₄ haloalkyl, or CN;    -   R⁵ is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl,        C₃₋₁₀ cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl —C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a2), SR^(a2),        C(O)R^(b2), C(O)NR^(c2)R^(d2), C(O)OR^(a2), OC(O)R^(b2),        OC(O)NR^(c2)R^(d2), NR^(c2)R^(d2) NR^(c2)C(O)R^(b2),        NR^(c2)C(O)OR^(a2), NR^(c2)C(O)NR^(c2)R^(d2), C(═NR^(e2))R^(b2),        C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)C(═NR^(e2))NR^(c2)R^(d2),        NR^(c2)S(O)R^(b2), NR^(c2)S(O)₂R^(b2),        NR^(c2)S(O)₂NR^(c2)R^(d2), S(O)R^(b2), S(O)NR^(c2)R^(d2),        S(O)₂R^(b2), and S(O)₂NR^(c2)R^(d2); wherein said C₁₋₆ alkyl,        C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl of R¹ are each optionally        substituted with 1, 2, 3, 4, or 5 substituents independently        selected from Cy³, Cy³-C₁₋₄ alkyl, halo, C₁₋₆ alkyl, C₂₋₆        alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a2),        SR^(a2), C(O)R^(b2), C(O)NR^(c2)R^(d2), C(O)OR^(a2),        OC(O)R^(b2), OC(O)NR^(c2)R^(d2), NR^(c2)R^(d2),        NR^(c2)C(O)R^(b2), NR^(c2)C(O)OR^(a2), NR^(c2)C(O)NR^(c2)R^(d2),        C(═NR^(e2))R^(b2), C(═NR^(e2))NR^(c2)R^(d2),        NR^(c2)C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)S(O)R^(b2),        NR^(c2)S(O)₂R^(b2), NR^(c2)S(O)₂NR^(c2)R^(d2), S(O)R^(b2),        S(O)NR^(c2)R^(d2), S(O)₂R^(b2), or S(O)₂NR^(c2)R^(d2);    -   each R⁶ is independently selected from H, halo, C₁₋₆ alkyl, C₁₋₆        haloalkyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl,        4-7 membered heterocycloalkyl, CN, NO₂, OR^(a3), SR^(a3),        C(O)R^(b3), C(O)NR^(c3)R^(d3), C(O)OR^(a3), OC(O)R^(b3),        OC(O)NR^(c3)R^(d3), NR^(c3)R^(d3), NR^(c3)C(O)R^(b3),        NR^(c3)C(O)NR^(c3)R^(d3), NR^(c3)C(O)OR^(a3),        C(═NR^(e3))NR^(c3)R^(d3), NR^(c3)C(═NR^(e3))NR^(c3)R^(d3),        S(O)R^(b3), S(O)NR^(c3)R^(d3), S(O)₂R^(b3), NR^(c3)S(O)₂R^(b3),        NR^(c3)S(O)₂NR^(c3)R^(d3), and S(O)₂NR^(c3)R^(d3), wherein said        C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6        membered heteroaryl, 4-7 membered heterocycloalkyl of R⁶ are        each optionally substituted with 1, 2, 3, 4, or 5 substituents        independently selected from halo, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆        alkynyl, C₁₋₆ haloalkyl, CN, NO₂, OR^(a3), SR^(a3), C(O)R^(b3),        C(O)NR^(c3)R^(d3), C(O)OR^(a3), OC(O)R^(b3), OC(O)NR^(c3)R^(d3),        NR^(c3)R^(d3), NR^(c3)C(O)R^(b3), NR^(c3)C(O)NR^(c3)R^(d3),        NR^(c3)C(O)OR^(a3), C(═NR^(e3))NR^(c3)R^(d3),        NR^(c3)C(═NR^(e3))NR^(c3)R^(d3), S(O)R^(b3), S(O)NR^(c3)R^(d3),        S(O)₂R^(b3), NR^(c3)S(O)₂R^(b3), NR^(c3)S(O)₂NR^(c3)R^(d3), and        S(O)₂NR^(c3)R^(d3);    -   each Cy¹ is independently selected from C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered        heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or        5 substituents independently selected from halo, C₁₋₆ alkyl,        C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a), SR^(a), C(O)R^(b),        C(O)NR^(c)R^(d), C(O)OR^(a), OC(O)R^(b), OC(O)NR^(c)R^(d),        NR^(c)R^(d), NR^(c)C(O)R^(b), NR^(c)C(O)OR^(a),        NR^(c)C(O)NR^(c)R^(d), C(═NR^(e))R^(b), C(═NR^(e))NR^(c)R^(d),        NR^(c)C(═NR^(e))NR^(c)R^(d), NR^(c)S(O)R^(b), NR^(c)S(O)₂R^(b),        NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b), S(O)NR^(c)R^(d), S(O)₂R^(b),        and S(O)₂NR^(c)R^(d);    -   each Cy² is independently selected from C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered        heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or        5 substituents independently selected from halo, C₁₋₆ alkyl,        C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a1), SR^(a1),        C(O)R^(b1), C(O)NR^(c1)R^(d1), C(O)OR^(a1), OC(O)R^(b1),        OC(O)NR^(c1)R^(d1), NR^(c1)R^(d1), NR^(c1)C(O)R^(b1),        NR^(c1)C(O)OR^(a1), NR^(c1)C(O)NR^(c1)R^(d1), C(═NR^(e1))R^(b1),        C(═NR^(e1))NR^(c1)R^(d1), NR^(c1)C(═NR^(e1))NR^(c1)R^(d1),        NR^(c1)S(O)R^(b1), NR^(c1)S(O)₂R^(b1),        NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1), S(O)NR^(c1)R^(d1),        S(O)₂R^(b1), and S(O)₂NR^(c1)R^(d1);    -   each Cy³ is independently selected from C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, and 4-14 membered        heterocycloalkyl, each optionally substituted by 1, 2, 3, 4, or        5 substituents independently selected from halo, C₁₋₆ alkyl,        C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₆₋₁₀ aryl, C₃₋₁₀        cycloalkyl, 5-14 membered heteroaryl, 4-14 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, CN, NO₂, OR^(a2), SR^(a2),        C(O)R^(b2), C(O)NR^(c2)R^(d2), C(O)OR^(a2), OC(O)R^(b2),        OC(O)NR^(c2)R^(d2), NR^(c2)R^(d2), NR^(c2)C(O)R^(b2),        NR^(c2)C(O)OR^(a2), NR^(c2)C(O)NR^(c2)R^(d2), C(═NR^(e2))R^(b2),        C(═NR^(e2))NR^(c2)R^(d2), NR^(c2)C(═NR^(e2))NR^(c2)R^(d2),        NR^(c2)S(O)R^(b2), NR^(c2)S(O)₂R^(b2),        NR^(c2)S(O)₂NR^(c2)R^(d2), S(O)R^(b2), S(O)NR^(c2)R^(d2),        S(O)₂R^(b2), and S(O)₂NR^(c2)R^(d2);    -   each R^(a), R^(b), R^(c), R^(d), R^(a1), R^(b1), R^(c1), R^(d1),        R^(a2), R^(b2), R^(c2), R^(d2), R^(a3), R^(b3), R^(c3), and        R^(d3) is independently selected from H, C₁₋₆ alkyl, C₁₋₆        haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇        cycloalkyl, 5-10 membered heteroaryl, 4-10 membered        heterocycloalkyl, C₆₋₁₀ aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄        alkyl, 5-10 membered heteroaryl-C₁₋₄ alkyl, and 4-10 membered        heterocycloalkyl-C₁₋₄ alkyl, wherein said C₁₋₆ alkyl, C₂₋₆        alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-10        membered heteroaryl, 4-10 membered heterocycloalkyl, C₆₋₁₀        aryl-C₁₋₄ alkyl, C₃₋₇ cycloalkyl-C₁₋₄ alkyl, 5-10 membered        heteroaryl-C₁₋₄ alkyl, and 4-10 membered heterocycloalkyl-C₁₋₄        alkyl of R″, R^(b), R^(c), R^(d), R^(a1), R^(b1), R^(c1),        R^(d1), R^(a2), R^(b2), R^(c2), R^(d2), R^(a3), R^(b3), R^(c3),        or R^(d3) is optionally substituted with 1, 2, 3, 4, or 5        substituents independently selected from halo, C₁₋₄ alkyl, C 1-4        haloalkyl, C₁₋₆ haloalkyl, C2-6 alkenyl, C2-6 alkynyl, CN,        OR^(a4), SR^(a4), C(O)R^(m), C(O)NR^(c4)R^(d4), C(O)OR^(a4),        OC(O)R^(b4), OC(O)NR^(c4)R^(d4), NR^(c4)R^(d4),        NR^(c4)C(O)R^(m), NR^(c4)C(O)NR^(c4)R^(d4), NR^(c4)C(O)OR^(a4),        C(═NR^(e4))NR^(c4)R^(d4), NR^(c4)C(═NR^(e4))NR^(c4)R^(d4),        S(O)R^(b4), S(O)NR^(c4)R^(d4), S(O)₂R^(b4), NR^(c4) S(O)₂R^(b4),        NR^(c4)S(O)₂NR^(c4)R^(d4), and S(O)₂NR^(c4)R^(d4);    -   each R^(a4), R^(b4), R^(c4), and R^(d4) are independently        selected from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆        alkynyl, C₆₋₁₀ aryl, C₃₋₇ cycloalkyl, 5-6 membered heteroaryl,        4-7 membered heterocycloalkyl, wherein said C₁₋₆ alkyl, C₁₋₆        haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₆₋₁₀ aryl, C₃₋₇        cycloalkyl, 5-6 membered heteroaryl, and 4-7 membered        heterocycloalkyl are each optionally substituted with 1, 2, or 3        substituents independently selected from OH, CN, amino, halo,        C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, and C₁₋₆ haloalkoxy;    -   each R^(e), R^(e1), R^(e2), R^(e3), and R^(e4) is independently        selected from H, C₁₋₄ alkyl, and CN;    -   R_(PTM) is H; halogen (e.g., Cl or F); —CN; —OH; —NO₂; —NH₂;        optionally substituted linear or branched alkyl (e.g.,        optionally substituted linear or branched C1-C6 alkyl or        optionally substituted linear or branched C1-C4 alkyl or C1-C8        alkyl optionally substituted with OH); optionally substituted        cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl,        cyclohexyl); O-optionally substituted linear or branched C1-C4        alkyl; an optionally substituted C1-C4 alkynyl; an optionally        substituted C1-C4 alkyne; optionally substituted linear or        branched hydroxyalkyl (e.g., optionally substituted linear or        branched C1-C7 hydroxyalkyl); optionally substituted        alkylcycloalkyl (e.g., includes optionally substituted C1-C6        alkyl, optionally substituted C3-C10 cycloalkyl; or both);        optionally substituted alkyl-aryl (e.g., includes an optionally        substituted linear or branched C1-C6 alkyl, an optionally        substituted 5-10 member heteroaryl, or both); optionally        substituted alkyl-heteroaryl (e.g., includes an optionally        substituted linear or branched C1-C6 alkyl, an optionally        substituted 5-10 member heteroaryl, or both); optionally        substituted alky 1-heteroaryl (e.g., includes a C1-C6 alkyl, an        optionally substituted 5 or 6 member heteroaryl, optionally        substituted with a C1-C4 alkyl; the heteroaryl is selected from        oxazol-4-yl, 1,3,4-triazol-2-yl, and imidazole-1-yl; or        combination thereof); optionally substituted —NH—        alkyl-heteroaryl (e.g., an optionally substituted linear or        branched C1-C5 alkyl, an optionally substituted 5-8 member        heteroaryl, optionally substituted with a C1-C4 alkyl,        N—CH₂-pyrazol-4-yl, or a combination thereof); optionally        substituted alkoxy (e.g., an optionally substituted linear or        branched C1-C6 alkyl or —OCH₃); optionally substituted        O-heterocyclyl (e.g., includes an optionally substituted 3-12 or        4-7 member heterocyclyl; an optionally substituted        heterocycloalkyl; an optionally substituted C₃₋₁₂ monocyclic or        bicyclic heterocycloalkyl; optionally substituted with at least        one OH, C1-C5 alkyl (such as a methyl), ═O, NH₂, or a        combination thereof; or a combination thereof); optionally        substituted S-heterocyclyl (e.g., includes an optionally        substituted 4-7 member heterocyclyl; an optionally substituted        heterocycloalkyl; optionally substituted with at least one C1-C4        alkyl (such as a methyl), ═O, or a combination thereof; or a        combination thereof); optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —(CH₂)_(u)CO(CH₂)_(v)CH₃, —COCH₃, or —CH₂CH₂COCH₃,        wherein each u and v is independently selected from 1, 2, 3, 4        or 5); optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —O(CH₂)_(u)CO(CH₂)_(v)CH₃,        —O(CH₂)_(u)CH((CH₂)_(x)CH₃)(CH₂)_(w)CO(CH₂)_(v)CH₃, —O—CH₂COCH₃,        —O—CH₂COCH₂CH₃, —O—CH(CH₃)COCH₃, —OCH₂COCH₃, or —OCH₂(CH₃)COCH₃,        wherein each u, v, w, and x is independently selected from 1, 2,        3, 4 or 5); optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a),        —CONR_(PTM1a)R_(PTM2a), —CH₂CONR_(PTM1a)R_(PTM2a),        —CH₂CH₂CONR_(PTM1a)R_(PTM2a), —CONHCH₃, or —CH₂CONHCH₃, wherein        each u and v is independently selected from 1, 2, 3, 4 or 5);        optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —O(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a),        —O(CH₂)_(u)CH((CH₂)_(x)CH₃)(CH₂)_(w)CO(CH₂)_(v)        NR_(PTM1a)R_(PTM2a), —O—CH(CH₃)CONR_(PTM1a)R_(PTM2a),        —O—CH₂CONR_(PTM1a)R_(PTM2a), Or —OCH₂C(O)NHOCH₃, wherein each u,        v, w, and x is independently selected from 1, 2, 3, 4 or 5);        optionally substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —(CH₂)_(u)CHCH(CH₂)_(w)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a) or        —CHCHCONR_(PTM1a)R_(PTM2a), wherein each u, v, and w is        independently selected from 1, 2, 3, 4 or 5); optionally        substituted

-   -    (e.g., optionally substituted with a linear or branched C1-C4        alkyl; —NH—(CH₂)_(u)CO(CH₂)_(v)NR_(PTM1a)R_(PTM2a) or        —NH—CH₂CONR_(PTM1a)R_(PTM2a), wherein each u and v is        independently selected from 1, 2, 3, 4 or 5); fluoroalkoxy        (e.g., a mono-, bi- and/or tri-fluoroalkoxy); optionally        substituted monocyclic or bicyclic cycloalkyl (e.g., an        optionally substituted 3-12 member cycloalkyl; optionally        substituted with at least one of OH, ═O, linear or branched        C1-C6 alkyl (such as a methyl, ethyl, or butyl), or NH₂; or a        combination thereof); optionally substituted hydroxycycloalkyl;        optionally substituted aryl (e.g., an optionally substitute        C5-C10 aryl, an optionally substituted 5-7 member aryl;        optionally substituted with at least one halogen or C1-C3 alkyl        (e.g., methyl or ethyl); or a combination thereof), optionally        substituted heteroaryl (e.g., an optionally substituted 5-10 or        member heteroaryl, an optionally substituted 5-7 member        heteroaryl; an optionally substituted 5-member heteroaryl;        optionally substituted with at least one halogen or C1-C3 alkyl        (e.g., methyl or ethyl); or a combination thereof) optionally        linked to Q via a C or N-atom of the heteroaryl (e.g., at least        one of optionally linked to Q, optionally linked via an        optionally substituted —(CH₂)_(u)O(CH₂)_(v)O(CH₂)_(x)—, or a        combination thereof); optionally substituted monocyclic or        bicyclic heterocyclyl (e.g., an optionally substituted 3-12        member heterocyclyl; an C3-C12 monocyclic or bicyclic        heterocycloalkyl, azetidine1-yl, pyrrolidin-1-yl, piperidin-1yl,        piperazin-1-yl, or morpholin-4-yl, or homopiperazin-1-yl, each        optionally substituted with OH, a linear or branched C1-C5 alkyl        (a methyl, ethyl, or butyl group) or NH₂) optionally linked to Q        via a C or N atom of the heterocyclyl (e.g., at least one of        optionally linked to Q, optionally linked via an optionally        substituted —(CH₂)_(u)O(CH₂)_(v)O(CH₂)_(x)—, or both);    -   t₁ is selected from 1, 2, 3, 4, or 5;    -   each t₂ is independently is independently selected from 0, 1, 2,        3, 4, or 5;    -   R_(PTM1a) and R_(PTM2a) are independently H, optionally        substituted C1-C4 alkyl (e.g., a CH₃ or CH₂CH₃), optionally        substituted C1-C4 alkoxy (e.g., —OCH₂ or —CH₂CH₃), optionally        substituted CH₂OCH₃ or R_(PTM1a) and R_(PTM2a) are joined        together form an optionally substituted 3-10 member ring;    -   n is an integer from 0 to 10; and        -   of the PTM indicates the site of attachment with a chemical            linking group or a CLM; and            (c) the L is a bond or a chemical linking group that            covalently couples the CLM to the PTM.

Therapeutic Compositions

The present invention further provides pharmaceutical compositionscomprising therapeutically effective amounts of at least onebifunctional compound as described herein, in combination with apharmaceutically acceptable carrier, additive or excipient.

In an additional aspect, the description provides therapeuticcompositions comprising an effective amount of a compound as describedherein or salt form thereof, and a pharmaceutically acceptable carrier,additive or excipient, and optionally an additional bioactive agent. Thetherapeutic compositions effect targeted protein degradation in apatient or subject, for example, an animal such as a human, and can beused for treating or ameliorating disease states or conditions which aremodulated by degrading the target protein. In certain embodiments, thetherapeutic compositions as described herein may be used to effectuatethe degradation of protein for the treatment or amelioration ofLRRK2-mediated neurodegenerative disease, inflammatory diseases,autoimmune diseases or cancer. In certain additional embodiments, thedisease is Parkinson's disease, Parkinson disease with dementia,Parkinson's disease at risk syndrome, dementia with Lewy bodies, Lewybody variant of Alzheimer's disease, combined Parkinson's disease andAlzheimer's disease, multiple system atrophy, striatonigraldegeneration, olivopontocerebellar atrophy, and Shy-Drager syndrome.

In alternative aspects, the present disclosure relates to a method fortreating a disease state or ameliorating one or more symptoms of adisease or condition in a subject in need thereof by degrading the LRRK2protein comprising administering to said patient or subject an effectiveamount, e.g., a therapeutically effective amount, of at least onecompound as described herein, optionally in combination with apharmaceutically acceptable carrier, additive or excipient, andoptionally coadministered with an additional bioactive agent, whereinthe composition is effective for treating or ameliorating the disease ordisorder or one or more symptoms thereof in the subject. The methodaccording to the present disclosure may be used to treat certain diseasestates or conditions including neurodegenerative diseases, by virtue ofthe administration of effective amounts of at least one compounddescribed herein.

The present disclosure further includes pharmaceutical compositionscomprising a pharmaceutically acceptable salt, in particular, acid orbase addition salts of compounds as described herein. The acids whichare used to prepare the pharmaceutically acceptable acid addition saltsof the aforementioned compounds useful according to this aspect arethose which form non-toxic acid addition salts, i.e., salts containingpharmacologically acceptable anions, such as the hydrochloride,hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acidphosphate, acetate, lactate, citrate, acid citrate, tartrate,bitartrate, succinate, maleate, fumarate, gluconate, saccharate,benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate,p-toluenesulfonate and pamoate [i.e., 1,1′-methylene-bis-(2-hydroxy-3naphthoate)]salts, among numerous others.

Pharmaceutically acceptable base addition salts may also be used toproduce pharmaceutically acceptable salt forms of the compoundsaccording to the present disclosure. The chemical bases that may be usedas reagents to prepare pharmaceutically acceptable base salts of thepresent compounds are those that form non-toxic base salts with suchcompounds. Such non-toxic base salts include, but are not limited tothose derived from such pharmacologically acceptable cations such asalkali metal cations (e.g., potassium and sodium) and alkaline earthmetal cations (e.g., calcium, zinc and magnesium), ammonium orwater-soluble amine addition salts such asN-methylglucamine-(meglumine), and the lower alkanolammonium and otherbase salts of pharmaceutically acceptable organic amines, among others.

The compounds as described herein may, in accordance with thedisclosure, be administered in single or divided doses by the oral,parenteral or topical routes. Administration of the active compound mayrange from continuous (intravenous drip) to several oral administrationsper day (for example, Q.I.D.) and may include oral, topical, parenteral,intramuscular, intravenous, sub-cutaneous, transdermal (which mayinclude a penetration enhancement agent), buccal, sublingual, intranasal, intra ocular, intrathecal, and suppository administration, amongother routes of administration. Enteric coated oral tablets may also beused to enhance bioavailability of the compounds from an oral route ofadministration. The most effective dosage form will depend upon thepharmacokinetics of the particular agent chosen as well as the severityof disease in the patient. Administration of compounds according to thepresent disclosure as sprays, mists, or aerosols for intra-nasal,intra-tracheal or pulmonary administration may also be used. The presentdisclosure therefore also is directed to pharmaceutical compositionscomprising an effective amount of compound as described herein,optionally in combination with a pharmaceutically acceptable carrier,additive or excipient. Compounds according to the present disclosure maybe administered in immediate release, intermediate release or sustainedor controlled release forms. Sustained or controlled release forms arepreferably administered orally, but also in suppository and transdermalor other topical forms. Intramuscular injections in liposomal form or indepot formulation may also be used to control or sustain the release ofcompound at an injection site.

The compositions as described herein may be formulated in a conventionalmanner using one or more pharmaceutically acceptable carriers and mayalso be administered in controlled-release formulations.Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as prolaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The compositions as described herein may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

Sterile injectable forms of the compositions as described herein may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1, 3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such as Ph. Helv orsimilar alcohol.

The pharmaceutical compositions as described herein may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers which are commonly used includelactose and corn starch. Lubricating agents, such as magnesium stearate,are also typically added. For oral administration in a capsule form,useful diluents include lactose and dried corn starch. When aqueoussuspensions are required for oral use, the active ingredient may becombined with emulsifying and suspending agents. If desired, certainsweetening, flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions as described herein maybe administered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient, which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions as described herein may also beadministered topically. Suitable topical formulations are readilyprepared for each of these areas or organs. For topical applications,the pharmaceutical compositions may be formulated in a suitable ointmentcontaining the active component suspended or dissolved in one or morecarriers. Carriers for topical administration of the compounds of thisdisclosure include, but are not limited to, mineral oil, liquidpetrolatum, white petrolatum, propylene glycol, polyoxyethylene,polyoxypropylene compound, emulsifying wax and water. In certainpreferred aspects of the disclosure, the compounds may be coated onto astent which is to be surgically implanted into a patient in order toinhibit or reduce the likelihood of occlusion occurring in the stent inthe patient.

Alternatively, the pharmaceutical compositions can be formulated in asuitable lotion or cream containing the active components suspended ordissolved in one or more pharmaceutically acceptable carriers. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions as described herein may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The amount of active pharmaceutical ingredient in a pharmaceuticalcomposition as described herein that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thecondition of the subject and disease treated, as well as the particularmode of administration. Preferably, the compositions should beformulated to contain between about 0.05 milligram and about 750milligrams or more, more preferably about 1 milligram to about 600milligrams, and even more preferably about 10 milligrams to about 500milligrams of active ingredient, alone or in combination with anothercompound according to the present disclosure.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity and bioavailability of the specificcompound employed, the age, body weight, general health, sex, diet, timeof administration, rate of excretion, drug combination, and the judgmentof the treating physician and the severity of the particular disease orcondition being treated.

A patient or subject in need of therapy using compounds according to themethods described herein can be treated by administering to the patient(subject) an effective amount of the compound according to the presentdisclosure depending upon the pharmaceutically acceptable salt, solvateor polymorph, thereof optionally in a pharmaceutically acceptablecarrier or diluent, either alone, or in combination with another knowntherapeutic agent.

The active compound is combined with the pharmaceutically acceptablecarrier or diluent in an amount sufficient to deliver to a patient atherapeutically effective amount for the desired indication, withoutcausing serious toxic effects in the patient treated. A preferred doseof the active compound for all of the herein-mentioned conditions is inthe range from about 10 nanograms per kilograms (ng/kg) to 300milligrams per kilograms (mg/kg), preferably 0.1 to 100 mg/kg per day,more generally 0.5 to about 25 mg per kilogram body weight of therecipient/patient per day. A typical topical dosage will range from0.01-5% wt/wt in a suitable carrier.

The compound is conveniently administered in any suitable unit dosageform, including but not limited to a dosage form containing less than 1milligrams (mg), 1 mg to 3000 mg, or 5 mg to 500 mg of active ingredientper unit dosage form. An oral dosage of about 25 mg-250 mg is oftenconvenient.

The active ingredient is preferably administered to achieve peak plasmaconcentrations of the active compound of about 0.00001-30 millimole(mM), preferably about 0.1-30 micromole (μM). This may be achieved, forexample, by the intravenous injection of a solution or formulation ofthe active ingredient, optionally in saline, or an aqueous medium oradministered as a bolus of the active ingredient. Oral administrationmay also be appropriate to generate effective plasma concentrations ofactive agent.

The concentration of active compound in the drug composition will dependon absorption, distribution, inactivation, and excretion rates of thedrug as well as other factors known to those of skill in the art. It isto be noted that dosage values will also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions, and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed composition. The active ingredient may be administered atonce, or may be divided into a number of smaller doses to beadministered at varying intervals of time.

Oral compositions will generally include an inert diluent or an ediblecarrier. They may be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound or its prodrug derivative can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Pharmaceuticallycompatible binding agents, and/or adjuvant materials can be included aspart of the composition.

The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a bindersuch as microcrystalline cellulose, gum tragacanth or gelatin; anexcipient such as starch or lactose, a dispersing agent such as alginicacid, Primogel, or corn starch; a lubricant such as magnesium stearateor Sterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring. When the dosage unitform is a capsule, it can contain, in addition to material of the abovetype, a liquid carrier such as a fatty oil. In addition, dosage unitforms can contain various other materials which modify the physical formof the dosage unit, for example, coatings of sugar, shellac, or entericagents.

The active compound or pharmaceutically acceptable salt thereof can beadministered as a component of an elixir, suspension, syrup, wafer,chewing gum or the like. A syrup may contain, in addition to the activecompounds, sucrose as a sweetening agent and certain preservatives, dyesand colorings and flavors.

The active compound or pharmaceutically acceptable salts thereof canalso be mixed with other active materials that do not impair the desiredaction, or with materials that supplement the desired action, such asanti-cancer agents, as described herein among others. In certainpreferred aspects of the disclosure, one or more compounds according tothe present disclosure are coadministered with another bioactive agent,such as an anti-cancer agent or a wound healing agent, including anantibiotic, as otherwise described herein.

Solutions or suspensions used for parenteral, intradermal, subcutaneous,or topical application can include the following components: a sterilediluent such as water for injection, saline solution, fixed oils,polyethylene glycols, glycerine, propylene glycol or other syntheticsolvents; antibacterial agents such as benzyl alcohol or methylparabens; antioxidants such as ascorbic acid or sodium bisulfite;chelating agents such as ethylenediaminetetraacetic acid; buffers suchas acetates, citrates or phosphates and agents for the adjustment oftonicity such as sodium chloride or dextrose. The parental preparationcan be enclosed in ampoules, disposable syringes or multiple dose vialsmade of glass or plastic.

If administered intravenously, preferred carriers are physiologicalsaline or phosphate buffered saline (PBS).

In any aspect or embodiment described herein, the active compounds areprepared with carriers that will protect the compound against rapidelimination from the body, such as a controlled release formulation,including implants and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, poly anhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Methods for preparation of suchformulations will be apparent to those skilled in the art.

Liposomal suspensions may also be pharmaceutically acceptable carriers.These may be prepared according to methods known to those skilled in theart, for example, as described in U.S. Pat. No. 4,522,811 (which isincorporated herein by reference in its entirety). For example, liposomeformulations may be prepared by dissolving appropriate lipid(s) (such asstearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline,arachadoyl phosphatidyl choline, and cholesterol) in an inorganicsolvent that is then evaporated, leaving behind a thin film of driedlipid on the surface of the container. An aqueous solution of the activecompound are then introduced into the container. The container is thenswirled by hand to free lipid material from the sides of the containerand to disperse lipid aggregates, thereby forming the liposomalsuspension.

Therapeutic Methods

In an additional aspect, the description provides therapeutic methodscomprising administration of an effective amount of a compound asdescribed herein or salt form thereof, and a pharmaceutically acceptablecarrier. The therapeutic methods are useful to effect proteindegradation in a patient or subject in need thereof, for example, ananimal such as a human, for treating or ameliorating a disease state,condition or related symptom that me be treated through targeted proteindegradation.

The terms “treat”, “treating”, and “treatment”, etc., as used herein,refer to any action providing a benefit to a patient for which thepresent compounds may be administered, including the treatment of anydisease state, condition, or symptom which is related to the protein towhich the present compounds bind. Disease states or conditions,including cancer, which may be treated using compounds according to thepresent disclosure are set forth hereinabove.

The description provides therapeutic methods for effectuating thedegradation of proteins of interest for the treatment or amelioration ofa disease, e.g., cancer. In any aspect or embodiment described herein,the disease is multiple myeloma. As such, in another aspect, thedescription provides a method of ubiquitinating/degrading a targetprotein in a cell. In certain embodiments, the method comprisesadministering a bifunctional compound of the invention. The control orreduction of specific protein levels in cells of a subject as affordedby the present disclosure provides treatment of a disease state,condition, or symptom. In any aspect or embodiment described herein, themethod comprises administering an effective amount of a compound asdescribed herein, optionally including a pharmaceutically acceptableexcipient, carrier, adjuvant, another bioactive agent or combinationthereof.

In additional embodiments, the description provides methods for treatingor ameliorating a disease, disorder or symptom thereof in a subject or apatient, e.g., an animal such as a human, comprising administering to asubject in need thereof a composition comprising an effective amount,e.g., a therapeutically effective amount, of a compound as describedherein or salt form thereof, and a pharmaceutically acceptableexcipient, carrier, adjuvant, another bioactive agent or combinationthereof, wherein the composition is effective for treating orameliorating the disease or disorder or symptom thereof in the subject.

In another aspect, the description provides methods for identifying theeffects of the degradation of proteins of interest in a biologicalsystem using compounds according to the present disclosure.

In another aspect, the description provides a process for making amolecule that can cause degradation of LRRK2 in a cell, comprising thesteps of: i. providing a small molecule that binds LRRK2; ii. providingand E3 ubiquitin ligase binding moiety (ULM), preferably a CLM such asthalidomide, pomalidomide, lenalidomide or an analog thereof; and iii.covalently coupling the small molecule of step (i) to the ULM of step(ii) via a chemical linking group (L) to form a compound which binds toboth a cereblon E3 ubiquitin ligase and LRRK2 protein in the cell, suchthat the cereblon E3 ubiquitin ligase is in proximity to, andubiquitinates LRRK2 protein bound thereto, such that the ubiquitinatedLRRK2 is then degraded.

In another aspect, the description provides a method for detectingwhether a molecule can trigger degradation of a LRRK2 protein in a cell,the method comprising the steps of: (i) providing a molecule for whichthe ability to trigger degradation of LRRK2 protein in a cell is to bedetected, said molecule comprising the structure: CLM-L-PTM, wherein CLMis a cereblon E3 ubiquitin ligase binding moiety capable of binding acereblon E3 ubiquitin ligase in a cell, which CLM is thalidomide,pomalidomide, lenalidomide, or an analog thereof; PTM is a proteintargeting moiety, which is a small molecule that binds to LRRK2, saidLRRK2 having at least one lysine residue available to be ubiquitinatedby a cereblon E3 ubiquitin ligase bound to the CLM of the molecule; andL is a chemical linking group that covalently links the CLM to the PTMto form the molecule; (ii) incubating a LRRK2 protein-expressing cell inthe presence of the molecule of step (i); and (iii) detecting whetherthe LRRK2 protein in the cell has been degraded.

In any of the aspects or embodiments described herein, the smallmolecule capable of binding LRRK2, is a small molecule as describedherein.

In another aspect of said treatment, the present disclosure provides amethod of treating a human patient in need of said treatment of adisease state, condition, or symptom causally related to LRRK2expression, over-expression, mutation, misfolding or dysregulation wherethe degradation of the LRRK2 protein will produce a therapeutic effectin the patient, the method comprising administering to the patient aneffective amount of a compound according to the present disclosure,optionally in combination with another bioactive agent. The diseasestate, condition, or symptom may be caused by a microbial agent or otherexogenous agent such as a vims, bacteria, fungus, protozoa or othermicrobe, or may be a disease state, which is caused by expression,overexpression, mutation, misfolding, or dysregulation of the protein,which leads to a disease state, condition, or symptom.

In another aspect, the present disclosure provides a method of treatingor ameliorating at least one symptom of a disease or condition in asubject, comprising the steps of: providing a subject identified ashaving a symptom of a disease or condition causally related toexpression, overexpression, mutation, misfolding, or dysregulation ofLRRK2 protein in the subject, and the symptom of the disease orcondition is treated or ameliorated by degrading LRRK2 protein in cellsof the subject; and administering to the subject therapeuticallyeffective amount of a compound comprising a small molecule of thepresent invention such that the LRRK2 protein is degraded, therebytreating or ameliorating at least one symptom of a disease or conditionin the subject.

The term “disease state or condition” is used to describe any diseasestate or condition wherein protein expression overexpression, mutation,misfolding, or dysregulation (e.g., the amount of protein expressed in apatient is elevated) occurs and where degradation of the LRRK2 proteinto reduce or stabilize the level of LRRK2 protein (whether mutated ornot) in a patient provides beneficial therapy or relief of symptoms to apatient in need thereof. In certain instances, the disease state,condition, or symptom may be cured.

Disease state, condition, or symptom which may be treated usingcompounds according to the present disclosure include, for example,inflammatory conditions, and immunological conditions, asthma,autoimmune diseases such as multiple sclerosis, neurodegenerativedisease such as Parkinson's disease, various cancers, ciliopathies,diabetes, heart disease, hypertension, inflammatory bowel disease,mental retardation, mood disorder, obesity, refractive error,infertility, Angelman syndrome, Canavan disease, Coeliac disease,Charcot-Marie-Tooth disease, Cystic fibrosis, Duchenne musculardystrophy, Haemochromatosis, Haemophilia, Klinefelter's syndrome,Neurofibromatosis, Phenylketonuria, Polycystic kidney disease, (PKD1) or4 (PKD2) Prader-Willi syndrome, Sickle-cell disease, Tay-Sachs disease,and Turner syndrome.

The term “neurodegenerative disease” (or Degenerative nerve diseases) isused throughout the specification to refer to the pathological processthat affect many of body's activities, such as balance, movement,talking, breathing, and heart function. Many of these diseases aregenetic, sometimes the cause is a medical condition such as alcoholism,a tumor, or a stroke. Other causes may include toxins, chemicals, andviruses, and sometimes the cause is unknown. Exemplary neurodegenerativedisease which may be treated by the present compounds either alone or incombination with at least one additional anti-neurodegenerative diseasetherapeutic agent include therapeutic agents to treat Parkinson'sdisease, Parkinson disease with dementia, Parkinson's disease at risksyndrome, dementia with Lewy bodies, Lewy body variant of Alzheimer'sdisease, combined Parkinson's disease and Alzheimer's disease, multiplesystem atrophy, striatonigral degeneration, olivopontocerebellaratrophy, Huntington's disease, multiple sclerosis, amyotrophic lateralsclerosis and Shy-Drager syndrome. For example, Carmustine, Bexarotene,Tamibarotene, Imatinib, Paclitaxel, azithromycin, erythromycin,Doxycycline, Rifampicin, acyclovir, penciclovir and foscamet.

The term “bioactive agent” is used to describe an agent, other than acompound according to the present disclosure, which is used incombination with a present compound as an agent with biological activityto assist in effecting an intended therapy, inhibition and/orprevention/prophylaxis for which the present compounds are used.Preferred bioactive agents for use herein include those agents whichhave pharmacological activity similar to that for which the presentcompounds are used or administered and include for example, anti-canceragents, antiviral agents, especially including anti-HIV agents andanti-HCV agents, antimicrobial agents, antifungal agents, etc.

The term “additional anti-neurodegenerative disease agent” is used todescribe an anti-neurodegenerative disease therapeutic agent, which maybe combined with a compound according to the present disclosure to treatneurodegenerative disease. These agents include, for example,Carmustine, Bexarotene, Tamibarotene, Imatinib, Paclitaxel,azithromycin, erythromycin, Doxycycline, Rifampicin, acyclovir,penciclovir and foscamet.

The term “pharmaceutically acceptable derivative” is used throughout thespecification to describe any pharmaceutically acceptable prodrug form(such as an ester, amide other prodrug group), which, uponadministration to a patient, provides directly or indirectly the presentcompound or an active metabolite of the present compound.

EXAMPLES Abbreviations

-   -   ACN Acetonitrile    -   AcOH Acetic acid    -   DCM Dichloromethane    -   DMF Dimethylformamide    -   DMSO Dimethyl Sulfoxide    -   DIPEA N, N-Diisopropylethylamine    -   EtOAc/EA Ethyl Acetate    -   EtOH Ethanol    -   HATU Hexafluorophosphate Azabenzotriazole Tetramethyl Uronium    -   HPLC High pressure liquid chromatography    -   Hz Hertz    -   KOAc Potassium acetate    -   LCMS Liquid Chromatography/Mass Spectrometry    -   MHz Megahertz    -   NMR Nuclear Magnetic Resonance    -   MeOH Methanol    -   MS Mass Spectrometry    -   PE Petroleum ether    -   Psi Pound-force per square inch    -   RT or r.t. Room temperature    -   TEA Triethylamine    -   THF Tetrahydrofuran    -   TFA Trifluoracetic acid    -   TLC Thin layer chromatography    -   TMS Trimethylsilyl

General Synthetic Approach

The synthetic realization and optimization of the bifunctional moleculesas described herein may be approached in a stepwise or modular fashion.For example, identification of compounds that bind to the targetprotein, i.e., LRRK2 can involve high or medium throughput screeningcampaigns if no suitable ligands are immediately available. It is notunusual for initial ligands to require iterative design and optimizationcycles to improve suboptimal aspects as identified by data from suitablein vitro and pharmacological and/or ADMET assays. Part of theoptimization/SAR campaign would be to probe positions of the ligand thatare tolerant of substitution and that might be suitable places on whichto attach the chemical linking group previously referred to herein.Where crystallographic or NMR structural data are available, these canbe used to focus such a synthetic effort.

In a very analogous way one can identify and optimize ligands for an E3Ligase.

With PTMs and ULMs (e.g. CLMs) in hand, one skilled in the art can useknown synthetic methods for their combination with or without a chemicallinking group(s). Chemical linking group(s) can be synthesized with arange of compositions, lengths and flexibility and functionalized suchthat the PTM and ULM groups can be attached sequentially to distal endsof the linker. Thus, a library of bifunctional molecules can be realizedand profiled in in vitro and in vivo pharmacological and ADMET/PKstudies. As with the PTM and ULM groups, the final bifunctionalmolecules can be subject to iterative design and optimization cycles inorder to identify molecules with desirable properties.

In some instances, protecting group strategies and/or functional groupinterconversions (FGIs) may be required to facilitate the preparation ofthe desired materials. Such chemical processes are well known to thesynthetic organic chemist and many of these may be found in texts suchas “Greene's Protective Groups in Organic Synthesis” Peter G. M. Wutsand Theodora W. Greene (Wiley), and “Organic Synthesis: TheDisconnection Approach” Stuart Warren and Paul Wyatt (Wiley).

Synthetic Procedures Synthesis of Intermediate 1:4,5-Dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid

Step 1: Ethyl5-methyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine-6-carboxylate

To a solution of 1H-tetrazol-5-amine (5 gram (g), 48.50 millimole(mmol), 1 equivalent (eq), H₂O) in water (100 milliliters (mL)) wasadded formaldehyde (5.46 g, 72.75 mmol, 5.01 mL, 1.5 eq) and ethylacetoacetate (6.31 g, 48.50 mmol, 6.13 mL, 1 eq). The mixture wasstirred at 100° C. for 9 hours (hr) to give white solution. Then thesolution became suspension. LCMS (EB16-543-P1A1) showed the reaction wascompleted. The mixture was cooled to 20° C. and concentrated in reducedpressure at 20° C. The suspension was filtered and the filter cake wasconcentrated in vacuum to give ethyl5-methyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine-6-carboxylate (7.2 g,34.42 mmol, 70.96% yield) as a white solid.

Step 2: Ethyl 4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylate

To a mixture of ethyl5-methyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine-6-carboxylate (7.2 g,34.42 mmol, 1 eq) and Cs₂CO₃ (12.33 g, 37.86 mmol, 1.1 eq) in MeCN (70mL) was added Mel (6.06 g, 42.68 mmol, 2.66 mL, 1.24 eq) in one portionat 20° C. under N₂. The mixture was stirred at 50° C. for 1 hour (h).LCMS (EB16-545-P1A1) showed the reaction was completed. The aqueousphase was extracted with ethyl acetate (50 mL*3). The combined organicphase was washed with brine (50 mL*2), dried with anhydrous Na₂SO₄,filtered and concentrated in vacuum to give ethyl4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylate (7.5 g, 33.60mmol, 97.61% yield) as a yellow solid.

Step 3: 4,5-Dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid

To a mixture of ethyl4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylate (4 g, 17.92mmol, 1 eq) in THF (30 mL) was added LiOH (1.29 g, 53.76 mmol, 3 eq) andH₂O (5 mL) in one portion at 20° C. under N₂. The mixture was stirred at60° C. for 16 hours. LCMS (EB16-548-P1A1) showed the reaction wascompleted. The mixture was cooled to 20° C. and concentrated in reducedpressure at 20° C. The residue was acidified to pH=1 with HCl. The solidformed and collected by filtration. The filter cake was filtered undervacuum to give 4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylicacid (2.9 g, 14.86 mmol, 82.92% yield) as a white solid.

Synthesis of Intermediate 2:4,5,7-Trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.Synthesis of Intermediate 2 was made in a manner analogous tointermediate 1 using acetaldehyde in place of formaldehyde in step 1.

Synthesis of Intermediate 3:4,5,7,7-Tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylic acid.Synthesis of Intermediate 3 was made in a manner analogous tointermediate 1 using ethyl 2-acetyl-3-methyl-but-2-enoate, prepared bythe condensation of acetone and ethyl acetoacetate, in step 1.

Exemplary Synthesis of Exemplary Compound 1:N-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamideStep 1: Tert-butyl(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazine-1-carboxylate

To a mixture of tert-butyl (3R,5S)-3,5-dimethylpiperazine-1-carboxylate(1 g, 4.67 mmol, 1 eq) and benzyl4-(p-tolylsulfonyloxymethyl)piperidine-1-carboxylate (1.88 g, 4.67 mmol,1 eq) in MeCN (5 mL) was added KI (2.32 g, 14.00 mmol, 3 eq) and DIPEA(1.81 g, 14.00 mmol, 2.44 mL, 3 eq) in one portion at 20° C. under N₂.The mixture was stirred at 100° C. for 16 hours. LCMS (EB16-606-P1A6)showed desired MS. The mixture was cooled to 20° C. and concentrated inreduced pressure at 20° C. The residue was poured into water (5 mL). Theaqueous phase was extracted with ethyl acetate (5 mL*3). The combinedorganic phase was washed with brine (5 mL*2), dried with anhydrousNa₂SO₄, filtered and concentrated in vacuum. The residue was purified bysilica gel chromatography (DCM:MeOH=10:1, Rf=0.59, 20 g, 0-14% (10 min)of Ethyl acetate in Petroleum ether, 14% (20 min) of Ethyl acetate inPetroleum ether) to give tert-butyl(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazine-1-carboxylate(1.22 g, 2.74 mmol, 58.67% yield) as a yellow gum.

Step 2: Benzyl4-[[(2R,6S)-2,6-dimethylpiperazin-1-yl]methyl]piperidine-1-carboxylate

To a solution of tert-butyl(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazine-1-carboxylate(1.22 g, 2.74 mmol, 1 eq) in DCM (5 mL) was added TFA (312.18 mg, 2.74mmol, 202.71 microliter (uL), 1 eq) in one portion at 20° C. under N₂.The mixture was stirred at 20° C. for 30 min. LCMS (EB16-610-P1A1)showed the reaction was completed. The residue was basified to pH=7-8with saturated NaHCO₃. The aqueous phase was extracted with DCM (5mL*3). The combined organic phase was washed with brine (5 mL*2), driedwith anhydrous Na₂SO₄, filtered and concentrated in vacuum to givebenzyl4-[[(2R,6S)-2,6-dimethylpiperazin-1-yl]methyl]piperidine-1-carboxylate(930 mg, 2.69 mmol, 98.32% yield) as a yellow gum.

Step 3: Benzyl4-[[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate

To a mixture of benzyl4-[[(2R,6S)-2,6-dimethylpiperazin-1-yl]methyl]piperidine-1-carboxylate(930 mg, 2.69 mmol, 1 eq) and 4-bromo-2-fluoro-pyridine (473.74 mg, 2.69mmol, 1 eq) in DMF (5 mL) was added Cs₂CO₃ (1.75 g, 5.38 mmol, 2 eq) inone portion at 25° C. The mixture was stirred at 100° C. for 12 hours.LCMS (EB16-612-P1A1) showed the reaction was completed. The mixture wascooled to 20° C. and concentrated in reduced pressure at 20° C. Theresidue was poured into water (5 mL). The aqueous phase was extractedwith ethyl acetate (5 mL*3). The combined organic phase was washed withbrine (5 mL*2), dried with anhydrous Na₂SO₄, filtered and concentratedin vacuum. The residue was purified by silica gel chromatography (12 g,0-50%(10 min) of Ethyl acetate in Petroleum ether, 50%(10 min) of Ethylacetate in Petroleum ether) to give benzyl4-[[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(580 mg, 1.16 mmol, 42.97% yield) as a yellow gum.

Step 4:[2-[(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]boronicacid

To a mixture of benzyl4-[[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(580 mg, 1.16 mmol, 1 eq)4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(440.57 mg, 1.73 mmol, 1.5 eq) and KOAc (340.53 mg, 3.47 mmol, 3 eq) indioxane (2 mL) was added Pd(dppf)Cl₂ (42.32 mg, 57.83 umol, 0.05 eq) oneportion at 25° C. under N₂. The mixture was stirred at 100° C. for 2hours. LCMS (EB16-615-P1A1) showed the reaction was completed. Themixture was cooled to 25° C., filtered and concentrated in vacuum togive[2-[(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]boronicacid (669 mg, crude) as a black brown solid. The crude product was usedinto the next step without further purification.

Step 5: Benzyl4-[[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate

To a mixture of[2-[(3R,5S)-4-[(1-benzyloxycarbonyl-4-piperidyl)methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]boronicacid (669 mg, 1.43 mmol, 1 eq), 3-bromo-5-nitro-1H-indazole (381.90 mg,1.58 mmol, 1.1 eq) and KOAc (422.34 mg, 4.30 mmol, 3 eq) in EtOH (10 mL)and H₂O (2 mL) was added 4-ditert-butylphosphanyl-N,N-dimethyl-aniline;dichloropalladium (101.57 mg, 143.45 umol, 101.57 uL, 0.1 eq) in oneportion at 25° C. under N₂. The mixture was stirred at 100° C. for 12hours. LCMS (EB16-616-P1A2) showed the reaction was completed. Themixture was cooled to 20° C. and concentrated in reduced pressure. Theresidue was poured into water (10 mL). The aqueous phase was extractedwith ethyl acetate (10 mL*3). The combined organic phase was washed withbrine (10 mL*2), dried with anhydrous Na₂SO₄, filtered and concentratedin vacuum. The residue was purified by silica gel chromatography (12 g,100-200 mesh silica gel, 0-100% (15 min) of Ethyl acetate in Petroleumether, 100% (15 min) of Ethyl acetate in Petroleum ether) to give benzyl4-[[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(609 mg, 980.78 umol, 68.37% yield, 94% purity) as a yellow gum.

Step 6: Benzyl4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate

To a mixture of benzyl4-[[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(609 mg, 1.04 mmol, 1 eq) and Fe (291.34 mg, 5.22 mmol, 5 eq) in H₂O (1mL) and EtOH (5 mL) was added NH₄Cl (279.06 mg, 5.22 mmol, 5 eq) in oneportion at 20° C. under N₂. The mixture was stirred at 80° C. for 1.5hours. LCMS (EB16-620-P1A1) showed the reaction was completed. Theresidue was filtered and solution was concentrated in vacuum to givebenzyl4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(550 mg, 814.53 umol, 78.07% yield, 82% purity) as a brown solid.

Step 7: Benzyl4-[[(2R,6S)-4-[4-[5-(tert-butoxycarbonylamino)-1H-indazol-3-yl]-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate

To a mixture of benzyl4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(400 mg, 722.42 umol, 1 eq) and Boc₂O (173.43 mg, 794.66 umol, 182.56uL, 1.1 eq) in THF (5 mL) was added DIPEA (140.05 mg, 1.08 mmol, 188.75uL, 1.5 eq) in one portion at 20° C. under N₂. The mixture was stirredat 20° C. for 16 hours. TLC showed the reaction was completed. Theresidue was poured into water (5 mL). The aqueous phase was extractedwith ethyl acetate (5 mL*3). The combined organic phase was washed withbrine (5 mL*2), dried with anhydrous Na₂SO₄, filtered and concentratedin vacuum. The residue was purified by silica gel chromatography (12 g,0-50% (10 min) of Ethyl acetate in Petroleum ether, 50% (10 min) ofEthyl acetate in Petroleum ether) to give benzyl4-[[(2R,6S)-4-[4-[5-(tert-butoxycarbonylamino)-1H-indazol-3-yl]-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(274 mg, 410.70 umol, 56.85% yield, 98% purity) as an off-white solid.

Step 8: Tert-butylN-[3-[2-[(3R,5S)-3,5-dimethyl-4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate

To a mixture of benzyl4-[[(2R,6S)-4-[4-[5-(tert-butoxycarbonylamino)-1H-indazol-3-yl]-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]piperidine-1-carboxylate(274 mg, 419.08 umol, 1 eq) in THF (5 mL) was added Pd/C (50 mg, 419.08umol, 10% purity, 1 eq) under N₂. The suspension was degassed undervacuum and purged with H₂ several times. The mixture was stirred underH₂ (15 psi) at 25° C. for 16 hours. TLC showed the reaction wascompleted. The reaction mixture was filtered with MeOH (20 mL*3) and thefilter was concentrated to give tert-butylN-[3-[2-[(3R,5S)-3,5-dimethyl-4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate(160 mg, 252.16 umol, 60.17% yield, 81.9% purity) as a white solid.

Step 9: Tert-butylN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate

To a mixture of tert-butylN-[3-[2-[(3R,5S)-3,5-dimethyl-4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate(160 mg, 307.88 umol, 1 eq) and1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperidine-4-carbaldehyde(170.58 mg, 461.82 umol, 1.5 eq) in MeOH (5 mL) was added AcOH (18.49mg, 307.88 umol, 17.61 uL, 1 eq) and borane; 2-methylpyridine (65.86 mg,615.76 umol, 2 eq) in one portion at 20° C. under N₂. The mixture wasstirred at 20° C. for 4 h. LCMS (EB16-636-P1A2) showed there was desiredMS. The residue was poured into water (10 mL). The aqueous phase wasextracted with ethyl acetate (10 mL*3). The combined organic phase waswashed with brine (10 mL*2), dried with anhydrous Na₂SO₄, filtered andconcentrated in vacuum. The residue was purified by silica gelchromatography (Dichloromethane:Methanol=10:1, Rf=0.09, 0-20% (15 min)of Methanol in Dichloromethane, 20% (5 min) of Methanol inDichloromethane) to give tert-butylN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate(220 mg, 231.83 umol, 75.30% yield, 92% purity) as a yellow gum.

Step 10:5-[4-[[4-[[(2R,6S)-4-[4-(5-Amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

To a mixture of tert-butylN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]carbamate(220 mg, 251.99 umol, 1 eq) in DCM (3 mL) was added TFA (86.20 mg,755.97 umol, 55.97 uL, 3 eq) in one portion at 20° C. under N₂. Themixture was stirred at 20° C. for 30 min. TLC (DCM:MeOH=3:1, Rf=0.14)showed the reaction was completed. The mixture was concentrated underreduced pressure to give5-[4-[[4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(300 mg, crude, TFA) as a yellow gum.

Step 11:N-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(Compound 1)

To a solution of 4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylicacid (Intermediate 1, 30.30 mg, 155.25 umol, 1.2 eq) and5-[4-[[4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(100 mg, 129.38 umol, 1 eq) in DMF (2 mL) was stirred at 0° C. for 10min, then was added DIPEA (83.61 mg, 646.88 umol, 112.68 uL, 5 eq) andHATU (49.19 mg, 129.38 umol, 1 eq). Then the mixture was stirred at 25°C. for 16 h under N₂. LCMS (EB16-642-P1A8) showed the reaction wascompleted. The residue was poured into water (10 mL). The aqueous phasewas extracted with ethyl acetate (10 mL*3). The combined organic phasewas washed with brine (10 mL*3), dried with anhydrous Na₂SO₄, filteredand concentrated in vacuum. The crude product was purified byreversed-phase HPLC (Column: Phenomenex luna C18 150*25 mm*10 um;Condition: water (0.2% FA)-ACN; Begin B: 0; End B: 40; FlowRate: 35mL/min; Gradient Time: 35 min; 100% B Hold Time: 4 min) to giveN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(29.3 mg, 30.53 umol, 23.60% yield, 99% purity) as a yellow solid.

¹H NMR: (400 MHz, DMSO-d₆) δ: 13.42 (s, 1H), 11.08 (s, 1H), 10.07 (s,1H), 8.67 (s, 1H), 8.22 (d, J=5.1 Hz, 1H), 7.67-7.51 (m, 3H), 7.28 (d,J=13.8 Hz, 2H), 7.23 (d, J=8.8 Hz, 1H), 7.16 (d, J=5.3 Hz, 1H), 5.30 (s,2H), 5.11-5.02 (m, 1H), 4.12 (d, J=11.1 Hz, 2H), 4.03 (d, J=12.9 Hz,2H), 3.44 (s, 3H), 3.00-2.80 (m, 5H), 2.72-2.53 (m, 6H), 2.39-2.30 (m,2H), 2.27 (s, 3H), 2.16 (d, J=5.8 Hz, 2H), 2.05-1.97 (m, 1H), 1.95-1.71(m, 7H), 1.37 (s, 1H), 1.12 (d, J=5.8 Hz, 10H).

Exemplary Synthesis of Exemplary Compound 2:N-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide

To a solution of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid (32.48mg, 155.25 umol, 1.2 eq) and5-[4-[[4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(100 mg, 129.38 umol, 1 eq) in DMF (2 mL) was stirred at 0° C. for 10min, then was added DIPEA (83.60 mg, 646.88 umol, 112.67 uL, 5 eq) andHATU (49.19 mg, 129.38 umol, 1 eq). Then the mixture was stirred at 25°C. for 16 h under N₂. LCMS (EB16-643-P1A1) showed the reaction wascompleted. The residue was poured into water (10 mL). The aqueous phasewas extracted with ethyl acetate (10 mL*3). The combined organic phasewas washed with brine (10 mL*3), dried with anhydrous Na₂SO₄, filteredand concentrated in vacuum. The crude product was purified byreversed-phase HPLC (Column: Phenomenex luna C18 150*25 mm*10 um;Condition: water (0.2% FA)-ACN; Begin B: 0; End B: 40; FlowRate: 35mL/min; Gradient Time: 35 min; 100% B Hold Time: 4 min) to giveN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(34.9 mg, 35.84 umol, 27.70% yield, 99% purity) as a yellow solid.

¹H NMR: (400 MHz, DMSO) δ: 13.42 (s, 1H), 11.08 (s, 1H), 10.28 (s, 1H),8.67 (s, 1H), 8.23 (d, J=5.3 Hz, 1H), 8.15 (s, 1H), 7.67-7.58 (m, 2H),7.56-7.50 (m, 1H), 7.28 (d, J=13.6 Hz, 2H), 7.23 (d, J=8.8 Hz, 1H), 7.16(d, J=5.4 Hz, 1H), 5.83-5.71 (m, 1H), 5.06 (dd, J=5.4, 12.9 Hz, 1H),4.12 (d, J=12.0 Hz, 2H), 4.03 (d, J=12.9 Hz, 2H), 3.44 (s, 3H),3.01-2.82 (m, 5H), 2.72-2.55 (m, 5H), 2.40-2.33 (m, 2H), 2.20 (s, 3H),2.17 (s, 2H), 2.06-1.97 (m, 1H), 1.94-1.71 (m, 7H), 1.56 (d, J=6.4 Hz,3H), 1.38 (s, 1H), 1.11 (d, J=5.8 Hz, 10H).

Exemplary Synthesis of Exemplary Compound 3:N-[3-[2-[(3S,5R)-4-[2-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethoxy]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamideStep 1:(2R,6S)-4-(4-bromo-2-pyridyl)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-piperazine

To a solution of (3R,5S)-1-(4-bromo-2-pyridyl)-3,5-dimethyl-piperazine(2 g, 7.40 mmol, 1 eq) and 2-(2,2-dimethoxyethoxy)ethyl4-methylbenzenesulfonate (2.3 g, 7.56 mmol, 1.02 eq) in CH₃CN (20 mL)was added DIEA (1.91 g, 14.81 mmol, 2.58 mL, 2 eq) and KI (2.46 g, 14.81mmol, 2 eq). After addition, the reaction mixture was stirred at 120° C.for 16 h. LCMS (EB12-469-P1B) showed desired MS. TLC(dichloromethane:methanol=10:1) showed several new spots. After cooling,the reaction mixture was filtered and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(0 to 100% ethyl acetate in petroleum ether) to afford(2R,6S)-4-(4-bromo-2-pyridyl)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-piperazine(2.1 g, 4.79 mmol, 64.73% yield, 91.8% purity) as a yellow oil.

Step 2:(2R,6S)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazine

To a solution of(2R,6S)-4-(4-bromo-2-pyridyl)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-piperazine(2.1 g, 5.22 mmol, 1 eq) and4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(1.72 g, 6.79 mmol, 1.3 eq) in dioxane (40 mL) was added KOAc (1.54 g,15.66 mmol, 3 eq) and Pd(dppf)Cl₂ (381.93 mg, 521.97 umol, 0.1 eq).After addition, the reaction mixture was stirred at 100° C. for 1 hunder N₂. LCMS (EB12-470-P1B1) showed desired MS. The reaction mixturewas filtered and concentrated under reduced pressure to afford(2R,6S)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazine(3.8 g, 4.98 mmol, 95.40% yield) as a brown oil.

Step 3:3-[2-[(3R,5S)-4-[2-(2,2-dimethoxyethoxy)ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole

A mixture of(2R,6S)-1-[2-(2,2-dimethoxyethoxy)ethyl]-2,6-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazine(3.8 g, 4.98 mmol, 1 eq), 3-bromo-5-nitro-1H-indazole (1.81 g, 7.47mmol, 1.5 eq), 4-ditert-butylphosphanyl-N,N-dimethyl-aniline;dichloropalladium (352.66 mg, 498.05 umol, 352.66 uL, 0.1 eq) and KOAc(1.47 g, 14.94 mmol, 3 eq) in EtOH (30 mL) and H₂O (6 mL) was stirred at100° C. for 12 h under N₂. LCMS (EB12-473-P1B) showed desired MS. Thereaction mixture was filtered and the filtrate was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (0 to 15% methanol in dichloromethane) to afford3-[2-[(3R,5S)-4-[2-(2,2-dimethoxyethoxy)ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(2 g, 3.80 mmol, 76.24% yield, 92% purity) as a brown gum.

Step 4:2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]acetaldehyde

To a solution of3-[2-[(3R,5S)-4-[2-(2,2-dimethoxyethoxy)ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(700 mg, 1.44 mmol, 1 eq) in THF (5 mL) was added H₂SO₄ (2 M, 4 mL)After addition, the reaction solution was stirred at 70° C. for 1 h. TLC(dichloromethane:methanol=10:1) showed starting material consumed and anew spot formed. After cooling, the reaction was diluted with water (10mL) and washed with ethyl acetate (2×10 mL). The aqueous phase wasraised to pH=14 with NaOH. The solid was collected by filtration. Thesolid was dried under reduced pressure to afford2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]acetaldehyde(500 mg, 1.04 mmol, 71.83% yield, 91% purity) as a yellow solid. Thecrude product was used for next step directly.

Step 5:5-[4-[2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

To a solution of2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]acetaldehyde(250 mg, 570.15 umol, 1 eq) and AcOH (525.00 mg, 8.74 mmol, 0.5 mL,15.33 eq) in DCE (10 mL) MeOH (10 mL) and DMSO (2 mL) was added2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-yl-isoindoline-1,3-dione (390.30mg, 1.14 mmol, 2.00 eq) and NaOAc (140.32 mg, 1.71 mmol, 3 eq). Thereaction solution was stirred at 25° C. for 30 min. Then NaBH₃CN (107.49mg, 1.71 mmol, 3 eq) was added and the reaction was stirred at 25° C.for 12 h. LCMS (EB12-480-P1D2) showed desired MS. TLC(dichloromethane:methanol=10:1) showed several new spots. The reactionsolution was diluted with water (20 mL) and extracted with ethyl acetate(3×20 mL). The organic layer was dried over sodium sulfate andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (0 to 15% methanol in chloromethane) to afford5-[4-[2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(250 mg, 294.18 umol, 51.60% yield, 90% purity) as a yellow solid.

Step 6:5-[4-[2-[2-[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

To a mixture of5-[4-[2-[2-[(2R,6S)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(250 mg, 326.87 umol, 1 eq) in EtOH (3 mL) and H₂O (0.5 mL) was added Fe(91.27 mg, 1.63 mmol, 5 eq) and NH₄Cl (87.42 mg, 1.63 mmol, 5 eq). Afteraddition, the reaction mixture was stirred at 80° C. for 1 h. LCMS(EB12-481-P1C1) showed desired MS. The reaction mixture was filtered andfiltrate was concentrated under reduced pressure. The resulting wasdissolved with 10% methanol in dichloromethane (10 mL). The mixture wasfiltered and filtrate was concentrated under reduced pressure to afford5-[4-[2-[2-[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(140 mg, 188.23 umol, 57.59% yield, 98.8% purity) as a yellow solid.

Step 7:N-[3-[2-[(3S,5R)-4-[2-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethoxy]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(Compound 3)

To a solution of5-[4-[2-[2-[(2S,6R)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethoxy]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(140 mg, 190.52 umol, 1 eq) and4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid (74.37 mg,381.03 umol, 2 eq) in DMF (2 mL) was added DIEA (123.11 mg, 952.58 umol,165.92 uL, 5 eq) and HATU (72.44 mg, 190.52 umol, 1 eq). After addition,the reaction solution was stirred at 25° C. for 12 h. LCMS(EB12-482-P1D) showed desired MS. The reaction solution was diluted withwater (10 mL) and extracted with dichloromethane (3×10 mL). The organiclayer was dried over sodium sulfate and concentrated under reducedpressure. The residue was purified by prep.HPLC (column: Phenomenex lunaC18 150*25 mm*10 um; mobile phase: [water (0.225% FA)-ACN]; B %: 0%-40%;35 min) to affordN-[3-[2-[(3S,5R)-4-[2-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethoxy]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(5.6 mg, 5.83 umol, 3.06% yield, 95% purity) as a yellow solid.

¹H NMR: (400 MHz, CD₃OD) δ: 8.65 (s, 1H), 8.40 (s, 1H), 8.17 (d, J=5.3Hz, 1H), 7.52 (d, J=8.9 Hz, 1H), 7.38-7.32 (m, 2H), 7.24 (d, J=5.3 Hz,1H), 7.19 (d, J=8.5 Hz, 1H), 6.93 (d, J=2.1 Hz, 1H), 6.75 (dd, J=2.1,8.5 Hz, 1H), 5.30 (s, 2H), 5.02 (br dd, J=5.5, 12.8 Hz, 1H), 4.53 (br t,J=11.4 Hz, 2H), 3.92-3.81 (m, 4H), 3.67 (br t, J=4.8 Hz, 2H), 3.55 (brs, 2H), 3.51 (s, 3H), 3.26-3.20 (m, 4H), 3.04-2.94 (m, 2H), 2.92-2.81(m, 1H), 2.76 (br d, J=2.8 Hz, 1H), 2.68-2.56 (m, 7H), 2.36 (s, 3H),2.14-2.05 (m, 1H), 1.48 (br d, J=5.4 Hz, 6H)

Exemplary Synthesis of Exemplary Compound 4:N-[3-[2-[(3S,5R)-4-[2-[2-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethoxy]ethoxy]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide

Compound 4 was prepared in a manner analogous to example 3.

¹H NMR: (400 MHz, MeOD) δ: 8.71 (s, 1H), 8.27 (d, J=5.4 Hz, 1H),7.55-7.48 (m, 2H), 7.41 (d, J=8.5 Hz, 1H), 7.38-7.31 (m, 2H), 6.99 (d,J=1.8 Hz, 1H), 6.87 (d, J=8.5 Hz, 1H), 5.29 (s, 2H), 5.05 (dd, J=5.4,12.7 Hz, 1H), 4.55 (d, J=13.9 Hz, 2H), 3.85 (d, J=4.5 Hz, 2H), 3.79-3.53(m, 10H), 3.51 (s, 3H), 3.22 (s, 4H), 3.05-2.93 (m, 2H), 2.91-2.51 (m,9H), 2.34 (s, 3H), 2.16-2.07 (m, 1H), 1.49 (d, J=6.3 Hz, 6H).

Exemplary Synthesis of Exemplary Compound 5:N-[3-[2-[(3S,5R)-4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamideStep 1: Tert-butyl4-[2-[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]ethyl]piperazine-1-carboxylate

To a solution of (3R,5S)-1-(4-bromo-2-pyridyl)-3,5-dimethyl-piperazine(1.5 g, 5.55 mmol, 1 eq) and tert-butyl4-(2-chloroethyl)piperazine-1-carboxylate (2.07 g, 8.33 mmol, 3.58 mL,1.5 eq) in NMP (20 mL) then was added Cs₂CO₃ (5.43 g, 16.66 mmol, 3 eq)and KI (4.61 g, 27.76 mmol, 5.0 eq). Then the mixture was stirred at140° C. for 16 h. TLC (Dichloromethane:Methanol=10:1, Rf=0.5) showed thereaction a new spot. The residue was diluted with H₂O (50 mL) extractedwith ethyl acetate (50 mL×3). The combined organic layers were washedwith brine (50 mL×2), dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by silica gel column chromatography (0 to 10% Methanol inDichloromethane) to give tert-butyl4-[2-[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]ethyl]piperazine-1-carboxylate(1.2 g, 2.26 mmol, 40.77% yield, 91% purity) as a yellow gum.

Step 2: afford tert-butyl4-[2-[(2R,6S)-2,6-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazine-1-carboxylate

To a solution of tert-butyl4-[2-[(2R,6S)-4-(4-bromo-2-pyridyl)-2,6-dimethyl-piperazin-1-yl]ethyl]piperazine-1-carboxylate(1.2 g, 2.49 mmol, 1 eq) and4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(1.26 g, 4.97 mmol, 2. eq) in dioxane (10 mL) was added KOAc (732.30 mg,7.46 mmol, 3 eq), Pd(dppf)Cl₂ (182.00 mg, 248.73 umol, 0.1 eq). Then themixture was stirred at 100° C. for 1 hr under N₂. TLC(Dichloromethane:Methanol=10:1, Rf=0.3) showed the reaction a new spot.The reaction mixture was filtered and concentrated under reducedpressure to afford tert-butyl4-[2-[(2R,6S)-2,6-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazine-1-carboxylate(1.3 g, crude) as a brown gum.

Step 3: Tert-butyl4-(2-((2R,6S)-2,6-dimethyl-4-(4-(5-nitro-1H-indazol-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperazine-1-carboxylate

To a solution of tert-butyl4-(2-((2R,6S)-2,6-dimethyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperazine-1-carboxylate(1.3 g, 2.46 mmol, 1 eq) and 3-bromo-5-nitro-1H-indazole (713.03 mg,2.95 mmol, 1.2 eq) in EtOH (10 mL) and H₂O (2 mL) was added KOAc (722.83mg, 7.37 mmol, 3 eq), 4-ditert-butylphosphanyl-N,N-dimethyl-aniline;dichloropalladium (173.84 mg, 245.50 umol, 173.84 uL, 0.1 eq). Then themixture was stirred at 100° C. for 2 hr under N₂. TLC(Dichloromethane:Methanol=10:1, Rf=0.4) showed the reaction a new spot.The reaction mixture was poured into H₂O (10 mL). The mixture wasextracted with ethyl acetate (15 mL*3). The organic phase was washedwith brine (10 mL), dried over anhydrous Na₂SO₄, concentrated in vacuumto give a residue. The residue was purified by silica gel columnchromatography (0 to 15% Methanol in Dichloromethane) to give tert-butyl4-(2-((2R,6S)-2,6-dimethyl-4-(4-(5-nitro-1H-indazol-3-yl)pyridin-2-yl)piperazin-1-yl)ethyl)piperazine-1-carboxylate(750 mg, 1.01 mmol, 41.12% yield, 76% purity) as a yellow solid.

Step 4:3-[2-[(3S,5R)-3,5-dimethyl-4-(2-piperazin-1-ylethyl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole

To a solution oftert-butyl4-[2-[(2S,6R)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazine-1-carboxylate(750 mg, 1.33 mmol, 1 eq) in DCM (10 mL) was added TFA (12.32 g, 108.05mmol, 8 mL, 81.35 eq). After addition, the reaction solution was stirredat 20° C. for 1 h. LCMS (EB12-483-P1B) showed desired MS. The reactionsolution was diluted with water (50 mL) and washed with dichloromethane(2×30 mL). The aqueous phase was basified to pH˜14 with NaOH andextracted with ethyl acetate (3×50 mL). The organic layer was dried oversodium sulfate and concentrated under reduced pressure to afford3-[2-[(3S,5R)-3,5-dimethyl-4-(2-piperazin-1-ylethyl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(450 mg, 852.42 umol, 64.18% yield, 88% purity) as a yellow solid.

Step 5:5-[4-[2-[(2S,6R)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

To a solution of3-[2-[(3S,5R)-3,5-dimethyl-4-(2-piperazin-1-ylethyl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(250 mg, 538.14 umol, 1 eq) and2-(2,6-dioxo-3-piperidyl)-5-fluoro-isoindoline-1,3-dione (222.97 mg,807.21 umol, 1.5 eq) in DMSO (3 mL) was added DIEA (208.65 mg, 1.61mmol, 281.20 uL, 3 eq). After addition, the reaction was stirred at 100°C. for 12 h. LCMS (EB12-486-P1B1) showed desired MS. TLC(dichloromethane:methanol=10:1) showed several new spots. After cooling,the reaction solution was diluted with water (10 mL) and extracted withdichloromethane (3×10 mL). The organic layer was washed with brine (2×15mL), dried over sodium sulfate and concentrated under reduced pressure.The residue was purified by silica gel column chromatography (0 to 10%methanol in dichloromethane) to afford5-[4-[2-[(2S,6R)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(150 mg, 168.57 umol, 31.32% yield, 81% purity) as a yellow solid.

Step 6:5-[4-[2-[(2S,6R)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

To a mixture of5-[4-[2-[(2S,6R)-2,6-dimethyl-4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(150 mg, 208.11 umol, 1 eq) in EtOH (3 mL) and H₂O (0.5 mL) was added Fe(58.11 mg, 1.04 mmol, 5 eq) and NH₄Cl (55.66 mg, 1.04 mmol, 5 eq). Afteraddition, the reaction was stirred at 80° C. for 1 h. LCMS(EB12-487-P1B1) showed the reaction was completed. After cooling, thereaction mixture was filtered and filtrate was concentrated underreduced pressure. The residue was triturated with 10% methanol inchloromethane at 20° C. for 15 min. The solid was removed by filtrationand filtrate was concentrated under reduced pressure to afford5-[4-[2-[(2S,6R)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(70 mg, 89.17 umol, 42.85% yield, 88% purity) as a yellow solid.

Step 7:N-[3-[2-[(3S,5R)-4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(Compound 5)

To a solution of5-[4-[2-[(2S,6R)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]ethyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(130 mg, 188.19 umol, 1 eq) and4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylicacid (37.14 mg,190.30 umol, 1.01 eq) in DMF (2 mL) was added DIEA (72.97 mg, 564.57umol, 98.34 uL, 3 eq) and HATU (71.56 mg, 188.19 umol, 1 eq). Afteraddition, the reaction solution was stirred at 25° C. for 18 h. LCMS(EB12-489-P1B72) showed desired MS. The reaction was diluted with water(5 mL) and extracted with ethyl acetate (3×5 mL). The organic layer wasdried over sodium sulfate and concentrated under reduced pressure. Theresidue was purified by prep.HPLC (column: PhenomenexLunaC18150*25 mm*10um; mobile phase: [water (0.225% FA)-ACN]; B %: 0-30%; 35 min) to affordN-[3-[2-[(3S,5R)-4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]ethyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5-dimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(10.7 mg, 12.26 umol, 6.51% yield, 99.42% purity) as a yellow solid.

¹H NMR: (400 MHz, METHANOL-d₄) δ: 8.69 (s, 1H), 8.41 (br s, 1H), 8.28(d, J=5.3 Hz, 1H), 7.54-7.47 (m, 2H), 7.45 (s, 1H), 7.36 (d, J=6.6 Hz,2H), 7.15 (s, 1H), 7.01 (br d, J=6.9 Hz, 1H), 5.29 (s, 2H), 5.08 (dd,J=5.5, 12.4 Hz, 1H), 4.59 (s, 3H), 4.43 (br s, 2H), 3.51 (s, 4H), 3.27(br s, 4H), 3.02 (br t, J=12.6 Hz, 2H), 2.93-2.82 (m, 1H), 2.80-2.75 (m,1H), 2.75-2.69 (m, 3H), 2.66 (br t, J=4.7 Hz, 4H), 2.35 (s, 3H),2.19-2.05 (m, 1H), 1.44 (br d, J=4.9 Hz, 6H)

Exemplary Synthesis of Exemplary Compound 6:N-(3-{2-[(3R,5S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}-3,5-dimethylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 6 was prepared in a manner analogous to ExemplaryCompound 1 using Intermediate 3.

Step 1

To a mixture of methyl 3-oxobutanoate (29 g, 249.75 mmol, 26.85 mL, 1eq), ZnCl2 (5.11 g, 37.46 mmol, 1.75 mL, 0.15 eq) and acetone (21.76 g,374.63 mmol, 27.54 mL, 1.5 eq) was added AC₂O (33.15 g, 324.68 mmol,30.41 mL, 1.3 eq) in one portion. The mixture was stirred at 50° C. for48 hours to give yellow solution. TLC (Petroleum ether: Ethylacetate=10:1, Rf=0.4) showed no start material and a new spot. Theresidue was diluted with H₂O (200 mL) extracted with ethyl acetate (200mL×3). The combined organic layers were washed with brine (75 mL), driedover anhydrous sodium sulfate, filtered and concentrated under reducedpressure to give a residue. The residue was purified by silica gelcolumn chromatography (0 to 10% Ethyl acetate in Petroleum ether) togive methyl 2-acetyl-3-methyl-but-2-enoate (18 g, 73.76 mmol, 29.53%yield, 64% purity) as a colorless oil.

Step 2

To a mixture of methyl 2-acetyl-3-methyl-but-2-enoate (18 g, 115.25mmol, 1 eq) and 1H-tetrazol-5-amine (9.80 g, 115.25 mmol, 1 eq) in EtOH(50 mL) and was added molecular sieves (10 g, 115.25 mmol, 1 eq) at 20°C. under N₂. The mixture was stirred at 50° C. for 16 hours to giveyellow solution. TLC (Petroleum ether: Ethyl acetate=10:1, Rf=0.1)showed no start material and a new spot. The reaction mixture wasfiltered and concentrated under reduced pressure. The crude product waspoured into H₂O (100 mL). The mixture was extracted with ethyl acetate(150 mL*3). The organic phase was washed with brine (80 mL), dried overanhydrous Na₂SO₄, concentrated in vacuum to give a residue. The residuewas purified by silica gel column chromatography (0 to 30% Ethyl acetatein Petroleum ether) to give methyl5,7,7-trimethyl-4H-tetrazolo[1,5-a]pyrimidine-6-carboxylate (11 g, 48.29mmol, 41.90% yield, 98% purity) as a white solid.

Step 3

To a mixture of methyl5,7,7-trimethyl-4H-tetrazolo[1,5-a]pyrimidine-6-carboxylate (11 g, 49.28mmol, 1 eq) in MeCN (100 mL) was added Mel (11.71 g, 82.50 mmol, 5.14mL, 1.67 eq) in one portion and Cs₂CO₃ (19.27 g, 59.13 mmol, 1.2 eq) at20° C. under N₂. The mixture was stirred at 50° C. for 1 hours to giveyellow solution. The residue was diluted with H₂O (200 mL) extractedwith ethyl acetate (100 mL×3). The combined organic layers were washedwith brine (85 mL), dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to give methyl4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylate (11 g,crude) as a yellow solid.

Step 4

To a mixture of methyl4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylate (640 mg,2.70 mmol, 1 eq) in THF (5 mL) was added LiOH (129.33 mg, 5.40 mmol, 2eq) in H₂O (5 mL) at 20° C. The mixture was stirred at 50° C. for 16hours to give yellow solution. TLC (Petroleum ether: Ethyl acetate=0:1,Rf=0.04) showed no start material. The THF was evaporated and the H₂Osolution was acidified for pH=3 with 1M HCl, extracted with EtOAc, driedover Na₂SO₄, filtered and concentrated under reduced pressure to give4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylic acid (424 mg,crude) as a white solid.

Step 5

To a mixture of5-[4-[[4-[[(2R,6S)-4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-dimethyl-piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(97 mg, 125.50 umol, 1 eq) and4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylic acid (45.68mg, 188.24 umol, 92% purity, 1.5 eq) in DMF (5 mL) was added DIEA (48.66mg, 376.49 umol, 65.58 uL, 3 eq) in one portion and HATU (47.72 mg,125.50 umol, 1 eq) at 25° C. under N₂. The mixture was stirred at 25° C.for 16 hours to give yellow solution. LCMS showed desired product. Theresidue was concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30mm*3 um; mobile phase: [water (0.1M FANH4)-ACN]; B %: 0%-30%, 40 min) toaffordN-[3-[2-[(3R,5S)-4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]-3,5-dimethyl-piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7,7-tetramethyl-tetrazolo[1,5-a]pyrimidine-6-carboxamide(36.1 mg, 36.18 umol, 28.83% yield, 98.04% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 8:(7R)—N-(3-{2-[(3S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 8 was prepared in a manner analogous to ExemplaryCompound 1 using enantiomer 2 of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.

Step 1

The product 4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylicacid (9.5 g, 45.41 mmol, 1 eq) was separated by SFC (Column: DAICELCHIRALPAK IC (250 mm*30 mm, 10 um); Condition: 0.1% NH₃H₂O ETOH; BeginB: 20; End B: 20; FlowRate: 100 mL/min) to give enantiomer 1 designatedas (7S)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(4.68 g, 22.15 mmol, 48.77% yield, 99% purity) (Rt=3.080 min, 4.68 g)and enantiomer 2 designated as(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(4.1 g, 19.40 mmol, 42.73% yield, 99% purity) (Rt=3.258 min, 4.1 g) bothas an off-white solid.

Enantiomer 1: Randomly assigned as(7S)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(4.68 g, 22.15 mmol, 48.77% yield, 99% purity) was obtained as anoff-white solid (analytical chiral HPLC: ee %=100%, 4.11 min, α=−211).

Enantiomer 2: Randomly assigned as(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(4.1 g, 19.40 mmol, 42.73% yield, 99% purity) was obtained as aoff-white solid (analytical chiral HPLC: ee %=100%, 4.68 min, α=266).

Exemplary Synthesis of Exemplary Compound 9:(7S)—N-(3-{2-[(3S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 9 was prepared in a manner analogous to ExemplaryCompound 8 using Enantiomer 1 of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.

Exemplary Synthesis of Exemplary Compound 10:N-(3-{2-[(3S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5-dimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 10 was prepared in a manner analogous to ExemplaryCompound 1 using Intermediate 1.

Exemplary Synthesis of Exemplary Compound 11:(7R)—N-{3-[2-(4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 11 was prepared in a manner analogous to ExemplaryCompound 1 using Enantiomer 2 of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.

Exemplary Synthesis of Exemplary Compound 12:(7S)—N-{3-[2-(4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 12 was prepared in a manner analogous to ExemplaryCompound 1 using Enantiomer 1 of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.

Exemplary Synthesis of Exemplary Compound 13:N-{3-[2-(4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5-dimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 13 was prepared in a manner analogous to ExemplaryCompound 1 using Intermediate 1.

Exemplary Synthesis of Exemplary Compound 14:(7R)—N-{3-[2-(4-{2-[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of 4-(2,2-dimethoxyethyl)piperidine (118.67 mg, 684.97umol, 1.2 eq) and[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl4-methylbenzenesulfonate (300 mg, 570.81 umol, 1 eq) and DIEA (737.71mg, 5.71 mmol, 994.22 uL, 10 eq) KI (947.54 mg, 5.71 mmol, 10 eq) inMeCN (10 mL). Then the mixture was stirred at 100° C. for 2 hours underN₂. The residue was diluted with H₂O 20 mL extracted with ethyl acetate(20 mL×3). The combined organic layers were washed with brine 15 mL,dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure to give a residue. The residue was purified by silicagel column chromatography (0 to 10% Methanol in Dichloromethane) to give5-[4-[[4-(2,2-dimethoxyethyl)-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(280 mg, 515.74 umol, 90.35% yield, 97% purity) as a yellow solid.

Step 2

To a solution of5-[4-[[4-(2,2-dimethoxyethyl)-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(280 mg, 531.69 umol, 1 eq) in THF (2 mL) was added HCl (2 M, 1.51 mL,5.69 eq) and stirred at 25° C. for 1 hour. The reaction mixture waspoured into H₂O (20 mL) and basified with aqueous NaHCO₃ till pH=8. Themixture was extracted with ethyl acetate (20 mL*5) and dried overanhydrous Na₂SO₄, concentrated in vacuum to give2-[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]acetaldehyde(250 mg, crude) as a yellow solid.

Step 3

To a solution of2-[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]acetaldehyde(250 mg, 520.23 umol, 1 eq) and5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole (178.57 mg, 550.57umol, 1.06 eq) in DCM (20 mL) and MeOH (30 mL) was added AcOH (3.12 mg,52.02 umol, 2.98 uL, 0.1 eq) and the mixture was stirred at 20° C. for20 minutes. Then the NaBH₃CN (98.07 mg, 1.56 mmol, 3 eq) was added ofthe solution and was stirred at 20° C. for 16 hours. TLC(Dichloromethane:Methanol=5:1, Rf=0.2) showed no start material and anew spot. The residue was concentrated under reduced pressure to give aresidue. The residue was purified by silica gel column chromatography (0to 50% Methanol in Dichloromethane) to2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-1-piperidyl]methyl]-1-piperidyl]isoindoline-1,3-dione(400 mg, 344.79 umol, 66.28% yield, 68% purity) as a yellow solid.

Step 4

To a solution of2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-1-piperidyl]methyl]-1-piperidyl]isoindoline-1,3-dione(400 mg, 507.04 umol, 1 eq) in EtOH (10. mL) and H₂O (5 mL) was addedNH₄Cl (135.61 mg, 2.54 mmol, 5 eq), Fe (141.58 mg, 2.54 mmol, 5 eq).Then the mixture was stirred at 90° C. for 1 hour under N₂. The reactionmixture was filtered and concentrated under reduced pressure. Theresidue was purified by prep-HPLC (Phenomenex luna C18 100*40 mm*3 um:water (0.05% NH3H2O+10 mM NH4HCO3)-ACN; B %: 18%-48%, 10 min) to afford5-[4-[[4-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(76 mg, 96.74 umol, 19.08% yield, 96.6% purity) as a yellow solid.

Step 5

To a solution of5-[4-[[4-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(45 mg, 59.30 umol, 1 eq) and(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(12.40 mg, 59.30 umol, 1 eq) in DMF (2 mL) was stirred at 20° C. for 10minutes, then was added DIEA (38.32 mg, 296.48 umol, 51.64 uL, 5 eq) andHATU (27.06 mg, 71.15 umol, 1.2 eq). Then the mixture was stirred at 70°C. for 3 hours under N₂. LCMS showed desired product MS. The residue waspurified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 um;mobile phase: water (0.225% FA)-ACN; B %: 0%-30%, 35 min) to afford(7R)—N-[3-[2-[4-[2-[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]ethyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(14.9 mg, 15.52 umol, 26.18% yield, 98.98% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 15:(7R)—N-(3-{2-[4-(3-{4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}propyl)piperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-yl-isoindoline-1,3-dione (500mg, 1.10 mmol, 1 eq, TFA) and 1-chloro-3-iodo-propane (1.12 g, 5.48mmol, 589.42 uL, 5 eq) in CH₃CN (5 mL) was added K₂CO₃ (151.42 mg, 1.10mmol, 1 eq). After addition, the reaction was stirred at 20° C. for 1hour. TLC showed there was a new spot. The residue was poured into water(5 mL) and stirred for 5 minutes. The aqueous phase was extracted withethyl acetate (5 mL*3). The combined organic phase was washed with brine(5 mL*2), dried with anhydrous Na₂SO₄, filtered and concentrated invacuum. The residue was purified by silica gel chromatography (4 g,Rf=0.43, 100-200 mesh silica gel, 0-70% (10 min) of Ethyl acetate inPetroleum ether, 70% (10 min) Ethyl acetate in Petroleum ether) to give5-[4-(3-chloropropyl)piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(260 mg, 564.85 umol, 51.56% yield, 91% purity) as a yellow gum.

Step 2

To a mixture of(7R)-4,5,7-trimethyl-N-[3-(2-piperazin-1-yl-4-pyridyl)-1H-indazol-5-yl]-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(55 mg, 113.28 umol, 1 eq) and5-[4-(3-chloropropyl)piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(47.45 mg, 113.28 umol, 1 eq) in MeCN (5 mL) was added KI (37.61 mg,226.55 umol, 2 eq) and DIPEA (29.28 mg, 226.55 umol, 39.46 uL, 2 eq) inone portion at 20° C. under N₂. The mixture was stirred at 80° C. for 10hours. LCMS showed there was ˜78% of desired MS. The mixture was cooledto 20° C. and concentrated in reduced pressure at 20° C. The residue waspoured into water (5 mL). The aqueous phase was extracted with ethylacetate (5 mL*3). The combined organic phase was washed with brine (5mL*2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum.The crude product was purified by reversed-phase HPLC (Column:3_Phenomenex Luna C18 75*30 mm*3 um; Condition: water (0.2% LA)-ACN;Begin B: 0; End B: 30; FlowRate: 25 mL/min; Gradient Time: 35 minutes;100% B Hold Time: 3 minutes) to give(7R)—N-[3-[2-[4-[3-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]propyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(19.5 mg, 21.34 umol, 18.84% yield, 95% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 17:(7R)—N-[3-(2-{4-[(1-{1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}azetidin-3-yl)methyl]piperazin-1-yl}pyridin-4-yl)-1H-indazol-5-yl]-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate(7.6 g, 40.59 mmol, 1 eq) in DCM (80 mL) was added HCl/MeOH (4 M, 38.00mL, 3.74 eq), the mixture was stirred at 25° C. for 2 hours. TLC(Petroleum ether: Ethyl acetate=1:2, KMn04, Plate1) showed a new spotformed and the starting materials consumed completely. The reactionmixture was concentrated under reduced pressure to remove DCM andHCl/MeOH to give a residue. The crude product was used into the nextstep without further purification. Compound azetidin-3-ylmethanol (5.7g, crude, HCl) was obtained as a light yellow liquid.

Step 2

To a solution of azetidin-3-ylmethanol (2 g, 16.18 mmol, 1 eq, HCl) andtert-butyl 4-oxopiperidine-1-carboxylate (4.84 g, 24.28 mmol, 1.5 eq) inDCM (10 mL) and MeOH (2 mL) was added AcONa (6.64 g, 80.92 mmol, 5 eq)and HOAc (1.94 g, 32.37 mmol, 1.85 mL, 2 eq) for 30 minutes, after themixture was added NaBH₃CN (3.05 g, 48.55 mmol, 3 eq). Then the mixturewas stirred 25° C. for 16 hours. TLC (Methanol:Dichloromethane=1:10, I₂,Plate1) showed a new spot formed and the starting materials wasremained. The reaction mixture was concentrated under reduced pressureto remove DCM. The residue was diluted with water 40 mL and extractedwith EA (30 mL*4). The combined organic layers were washed with brine(60 mL), dried over Na₂SO₄ filtered and concentrated under reducedpressure to give a residue. The residue was purified by flash silica gelchromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of0-10% Methanol/Dichloromethane ether gradient @60 mL/min) to affordtert-butyl 4-[3-(hydroxymethyl)azetidin-1-yl]piperidine-1-carboxylate(2.2 g, 8.14 mmol, 50.28% yield) was obtained as a colorless gum.

Step 3

To a solution of tert-butyl4-[3-(hydroxymethyl)azetidin-1-yl]piperidine-1-carboxylate (2.2 g, 8.14mmol, 1 eq) in MeOH (10 mL) was added HCl/dioxane (4 M, 10 mL). Afteraddition, the reaction was stirred at 20° C. for 2 hours. LCMS showeddesired mass (Rt=0.220 min). The reaction was concentrated under reducedpressure to afford [1-(4-piperidyl)azetidin-3-yl]methanol (1.8 g, crude,HCl) as a white solid. The crude product was used for next stepdirectly.

Step 4

To a solution of [1-(4-piperidyl)azetidin-3-yl]methanol (1.7 g, 8.22mmol, 1 eq, HCl) in DMSO (20 mL) was added2-(2,6-dioxo-3-piperidyl)-5-fluoro-isoindoline-1,3-dione (2.27 g, 8.22mmol, 1 eq) and DIEA (3.19 g, 24.67 mmol, 4.30 mL, 3 eq). Afteraddition, the reaction solution was stirred at 100° C. for 2 hours. LCMSshowed desired mass. After cooling, the reaction was diluted with water(100 mL) and extracted with 10% methanol in dichloromethane (5×150 mL).The organic layer was dried over Na₂SO₄ and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(0 to 20% methanol in dichloromethane) to afford2-(2,6-dioxo-3-piperidyl)-5-[4-[3-(hydroxymethyl)azetidin-1-yl]-1-piperidyl]isoindoline-1,3-dione(1.4 g, 3.05 mmol, 37.12% yield, 93% purity) as a yellow solid.

Step 5

To a solution of2-(2,6-dioxo-3-piperidyl)-5-[4-[3-(hydroxymethyl)azetidin-1-yl]-1-piperidyl]isoindoline-1,3-dione(1.2 g, 2.81 mmol, 1 eq) in DCM (15 mL) was added TosCl (1.07 g, 5.63mmol, 2 eq), DMAP (34.38 mg, 281.38 umol, 0.1 eq) and TEA (854.19 mg,8.44 mmol, 1.17 mL, 3 eq). After addition, the reaction was stirred at20° C. for 16 hours. LCMS showed desired MS. The reaction was dilutedwith water (30 mL) and extracted with dichloromethane (3×20 mL). Theorganic layer was dried over sodium sulfate and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (0 to 6% methanol in dichloromethane) to afford[1-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]azetidin-3-yl]methyl4-methylbenzenesulfonate (460 mg, 741.51 umol, 26.35% yield, 93.6%purity) as a yellow solid.

Step 6

To a solution of[1-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]azetidin-3-yl]methyl4-methylbenzenesulfonate (77 mg, 132.61 umol, 1.07 eq) and(7R)-4,5,7-trimethyl-N-[3-(2-piperazin-1-yl-4-pyridyl)-1H-indazol-5-yl]-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(60 mg, 123.57 umol, 1 eq) in CH₃CN (2 mL) was added KI (41.03 mg,247.15 umol, 2 eq) and DIEA (31.94 mg, 247.15 umol, 43.05 uL, 2 eq).After addition, the reaction was stirred at 80° C. for 16 hours. LCMSshowed desired mass. After cooling, the reaction mixture was filteredand the filter cake was washed with DMSO (2 mL). The filtrate waspurified by prep.HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 um;mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 30%-60%; 35 min) toafford(7R)—N-[3-[2-[4-[[1-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]azetidin-3-yl]methyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(9.5 mg, 10.50 umol, 8.50% yield, 98.81% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 18:(7R)—N-(3-{2-[4-({1′-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]-[1,4′-bipiperidin]-4-yl}methyl)piperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of 5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole (1g, 3.08 mmol, 1 eq) and tert-butyl 4-formylpiperidine-1-carboxylate(723.32 mg, 3.39 mmol, 1.1 eq) in MeOH (10 mL) and HOAc (1 mL) was addedborane; 2-methylpyridine (659.57 mg, 6.17 mmol, 2 eq). After addition,the reaction mixture was stirred at 30° C. for 2 hours. LCMS showeddesired mass. The reaction mixture was diluted with water (30 mL) andbasified to pH 12 with solid NaOH. Then the mixture was filtered toafford filtrate cake. The residue was purified by silica gel columnchromatography (0 to 10% methanol in dichloromethane) to affordtert-butyl4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(1.6 g, 2.99 mmol, 97.04% yield, 97.54% purity) as a yellow solid.

Step 2

To a solution of tert-butyl4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(1.6 g, 3.07 mmol, 1 eq) in MeOH (10 mL) was added HCl/dioxane (4 M, 10mL, 13.04 eq). After addition, the reaction solution was stirred at 20°C. for 16 hours. LCMS showed desired mass. The reaction was concentratedunder reduced pressure. The resulting was dissolved in water (50 mL) andneutralized to pH 7 with solid NaOH. The solid was collected byfiltration to afford5-nitro-3-[2-[4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(860 mg, crude) as a yellow solid. The crude product was used for nextstep directly.

Step 3

To a solution of5-nitro-3-[2-[4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(860 mg, 2.04 mmol, 1 eq) and tert-butyl 4-oxopiperidine-1-carboxylate(2.03 g, 10.20 mmol, 5 eq) in MeOH (50 mL) and HOAc (5 mL) was addedborane; 2-methylpyridine (654.72 mg, 6.12 mmol, 3 eq). After addition,the reaction solution was stirred at 35° C. for 12 hours. LCMS desiredMS. The reaction was diluted with water (100 mL) and washed with ethylacetate (100 mL). The aqueous phase was basified to pH 14 with solid KOHand extracted with ethyl acetate (3×50 mL). The aqueous phase wasfiltered to collect the solid. The organic layer was dried over sodiumsulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (0 to 50% methanol indichloromethane) to afford tert-butyl4-[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]piperidine-1-carboxylate(750 mg, 1.14 mmol, 55.88% yield, 91.94% purity) as a yellow solid.

Step 4

To a solution of tert-butyl4-[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]piperidine-1-carboxylate(750 mg, 1.24 mmol, 1 eq) in DCM (5 mL) was added TFA (3.08 g, 27.01mmol, 2 mL, 21.78 eq). After addition, the reaction was stirred at 20°C. for 1 hour. LCMS showed desired MS. The reaction mixture wasconcentrated under reduced pressure to afford5-nitro-3-[2-[4-[[1-(4-piperidyl)-4-piperidyl]methyl]piperazin-1-yl]-4-pyridyl]-1H-indazole(900 mg, crude, TFA) as a yellow gum. The crude product was used fornext step directly.

Step 5

To a solution of5-nitro-3-[2-[4-[[1-(4-piperidyl)-4-piperidyl]methyl]piperazin-1-yl]-4-pyridyl]-1H-indazole(900 mg, 1.45 mmol, 1 eq, TFA) and2-(2,6-dioxo-3-piperidyl)-5-fluoro-isoindoline-1,3-dione (602.76 mg,2.18 mmol, 1.5 eq) in DMSO (10 mL) was added DIEA (940.08 mg, 7.27 mmol,1.27 mL, 5 eq). After addition, the reaction was stirred at 100° C. for12 hours. LCMS showed desired MS. After cooling, the reaction solutionwas diluted with water (100 mL). The solid was collected by filtrationand dried under reduced pressure to afford2-(2,6-dioxo-3-piperidyl)-5-[4-[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]-1-piperidyl]isoindoline-1,3-dione(800 mg, 862.21 umol, 59.27% yield, 82% purity) as a yellow solid. Thecrude product was used for next step directly.

Step 6

To a mixture of2-(2,6-dioxo-3-piperidyl)-5-[4-[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]-1-piperidyl]isoindoline-1,3-dione(400 mg, 525.74 umol, 1 eq) in EtOH (3 mL) and H₂O (0.5 mL) was added Fe(146.80 mg, 2.63 mmol, 5 eq) and NH₄Cl (140.61 mg, 2.63 mmol, 5 eq).After addition, the reaction was stirred at 80° C. for 2 hours. LCMSshowed desired MS. After cooling, the reaction was filtered and filtratewas concentrated in vacuo. The resulting was dissolved in DMSO (10 mL)and then water (80 mL) was added. The solid was collected by filtration.The solid was dried under reduced pressure to afford5-[4-[4-[[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(130 mg, 125.40 umol, 23.85% yield, 70.5% purity) as a yellow solid.

Step 7

To a solution of(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(37.21 mg, 177.87 umol, 1 eq) and5-[4-[4-[[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(130 mg, 177.87 umol, 1 eq) in DMF (2 mL) was added DIEA (68.97 mg,533.62 umol, 92.94 uL, 3 eq) and HATU (67.63 mg, 177.87 umol, 1 eq).After addition, the reaction solution was stirred at 25° C. for 16hours. LCMS showed desired MS. The reaction was filtered and filtratecake was washed with DMSO (1 mL). The filtrate was purified by prep.HPLC(column: 3_Phenomenex Luna C18 75*30 mm*3 um; mobile phase: [water(0.225% FA)-ACN]; B %: 0-30%; 35 min) to afford(7R)—N-[3-[2-[4-[[1-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]-4-piperidyl]methyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(10.7 mg, 11.45 umol, 6.44% yield, 98.66% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 20:(7R)—N-[3-(2-{4-[2-(2-{6-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]-2,6-diazaspiro[3.3]heptan-2-yl}ethoxy)ethyl]piperazin-1-yl}pyridin-4-yl)-1H-indazol-5-yl]-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 20 was prepared in a manner analogous to ExemplaryCompound 3.

Exemplary Synthesis of Exemplary Compound 21:(7S)—N-{3-[2-(4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of 5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole (500mg, 1.54 mmol, 1 eq) and tert-butyl4-(2-chloroethyl)piperazine-1-carboxylate (500 mg, 2.01 mmol, 1.3 eq) inCH₃CN (10 mL) was added KI (511.82 mg, 3.08 mmol, 2 eq) and DIEA (398.48mg, 3.08 mmol, 537.04 uL, 2 eq). After addition, the reaction wasstirred at 80° C. for 3 hours. LCMS showed desired MS. After cooling,the reaction mixture was diluted with water (10 mL) and extracted withdichloromethane (3×10 mL). The organic layer was dried over sodiumsulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (0 to 10% methanol indichloromethane) to afford tert-butyl4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazine-1-carboxylate(303 mg, 558.99 umol, 36.26% yield, 99% purity) as a yellow solid.

Step 2

To a solution of tert-butyl4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazine-1-carboxylate(303 mg, 564.64 umol, 1 eq) in DCM (3 mL) was added TFA (770.00 mg, 6.75mmol, 0.5 mL, 11.96 eq). After addition, the reaction was stirred at 20°C. for 1 hour. LCMS showed reactant consumed and desired MS wasdetected. The reaction mixture was concentrated in vacuo. The resultingwas dissolved in DCM (2 mL) and treated with DIEA (1 mL). The mixturewas concentrated under reduced pressure to afford5-nitro-3-[2-[4-(2-piperazin-1-ylethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(240 mg, crude) as a yellow solid. The crude product was used for nextstep directly.

Step 3

To a solution of5-nitro-3-[2-[4-(2-piperazin-1-ylethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(240 mg, 549.82 umol, 1 eq) and1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperidine-4-carbaldehyde(203.09 mg, 549.82 umol, 1 eq) in MeOH (10 mL) and HOAc (1 mL) was addedborane; 2-methylpyridine (117.62 mg, 1.10 mmol, 2 eq). After addition,the reaction solution was stirred at 20° C. for 2 hour. LCMS showedstarting material consumed and desired MS was detected. The reaction wasdiluted with water (20 mL) and extracted with dichloromethane (3×15 mL).The organic layer was dried over sodium sulfate and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (0 to 40% methanol in dichloromethane) to afford2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]methyl]-1-piperidyl]isoindoline-1,3-dione(445 mg, 523.38 umol, 95.19% yield, 92.9% purity) as a yellow solid.

Step 4

To a solution of2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]methyl]-1-piperidyl]isoindoline-1,3-dione(445 mg, 563.38 umol, 1 eq) in EtOH (5 mL) and H2O (1 mL) was added Fe(157.31 mg, 2.82 mmol, 5 eq) and NH4Cl (150.68 mg, 2.82 mmol, 5 eq).After addition, the reaction was stirred at 80° C. for 1 hour. LCMSshowed desired MS. After cooling, the reaction mixture was filtered andfiltrate was concentrated in vacuo. The residue was triturated with 10%methanol in dichloromethane at 20° C. for 10 minutes. The resultingmixture was filtered and the filtrate was concentrated under reducedpressure to afford5-[4-[[4-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(120 mg, 138.97 umol, 24.67% yield, 88% purity) as a yellow solid.

Step 5

To a solution of5-[4-[[4-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]piperazin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(55 mg, 72.38 umol, 1 eq) and(7S)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid (17mg, 81.26 umol, 1.12 eq) in DMF (2 mF) was added DIEA (46.77 mg, 361.89umol, 63.03 uL, 5 eq) and HATU (27.52 mg, 72.38 umol, 1 eq). Afteraddition, the reaction was stirred at 25° C. for 12 hours. LCMS showeddesired MS. The reaction was diluted with water (10 mF) and extractedwith dichloromethane (3×15 mF). The organic layer was dried over sodiumsulfate and concentrated under reduced pressure. The residue waspurified by prep.HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 um;mobile phase: [water (0.225% FA)-ACN]; B %: 0-35%; 35 min) to afford(7S)—N-[3-[2-[4-[2-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]piperazin-1-yl]ethyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(12.8 mg, 13.11 umol, 18.11% yield, 97.40% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 22:(7R)—N-{3-[2-(4-{2-[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]-2,2-difluoroethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 22 was prepared in a manner analogous to ExemplaryCompound 1 using tert-butyl4-[2-[4-(4-bromo-2-pyridyl)piperazin-1-yl]-1,1-difluoro-ethyl]piperidine-1-carboxylate.

Step 1

To a solution of tert-butyl 4-(2-bromoacetyl)piperidine-1-carboxylate(2.78 g, 9.08 mmol, 1 eq) in MeCN (10 mL) was stirred at 20° C. Then themixture was added benzyl piperazine-1-carboxylate (2 g, 9.08 mmol, 1.75mL, 1 eq) and stirred at 20° C. for 16 hr under N₂. TLC(Dichloromethane:Methanol=10:1, Rf=0.6) showed no start material and anew spot. The residue was diluted with H₂O (200 mL) extracted with ethylacetate (70 mL×3). The combined organic layers were washed with brine(80 mL), dried over anhydrous sodium sulfate, filtered and concentratedunder reduced pressure to give a residue. The residue was purified bysilica gel column chromatography (0 to 10% Dichloromethane in Methanol)to give benzyl4-[2-(1-tert-butoxycarbonyl-4-piperidyl)-2-oxo-ethyl]piperazine-1-carboxylate(1.9 g, 3.84 mmol, 42.27% yield, 90% purity) as a yellow gum.

Step 2

To a solution of benzyl4-[2-(1-tert-butoxycarbonyl-4-piperidyl)-2-oxo-ethyl]piperazine-1-carboxylate(1.9 g, 4.26 mmol, 1 eq) in DCM (30 mL) was stirred at 0° C. for 20 min.Then the mixture was added DAST (24.06 g, 149.25 mmol, 19.72 mL, 35 eq)and stirred at 20° C. for 16 hours under N₂. TLC (Petroleum ether: Ethylacetate=1:1, Rf=0.5) showed no start material and a new spot. Thereaction was cooled to 0° C. and quenched with aqueous NaHCO₃ (200 mL)extracted with ethyl acetate (100 mL×2). The combined organic layerswere washed with brine (45 mL), dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by silica gel column chromatography (0 to 50% Ethylacetate in Petroleum ether) to give benzyl4-[2-(1-tert-butoxycarbonyl-4-piperidyl)-2,2-difluoro-ethyl]piperazine-1-carboxylate(1.4 g, 2.55 mmol, 59.68% yield, 85% purity) as a Colorless gum.

Step 3

To a solution of benzyl4-[2-(1-tert-butoxycarbonyl-4-piperidyl)-2,2-difluoro-ethyl]piperazine-1-carboxylate(1.4 g, 2.99 mmol, 1 eq) in EtOH (10 mL) and EtOAc (10 mL). Then themixture was added Pd/C (0.2 g, 299.43 umol, 10% purity, 0.1 eq) andstirred at 25° C. for 16 hours under H₂ (15 PSI). TLC (Petroleum ether:Ethyl acetate=1:1, Rf=0.01) showed no start material and a new spot. Thereaction was filtered and concentrated under reduced pressure to givetert-butyl4-(1,1-difluoro-2-piperazin-1-yl-ethyl)piperidine-1-carboxylate (900 mg,crude) as a colorless gum.

Step 4

To a solution of tert-butyl4-(1,1-difluoro-2-piperazin-1-yl-ethyl)piperidine-1-carboxylate (900 mg,2.70 mmol, 1 eq) and 4-bromo-2-fluoro-pyridine (500 mg, 2.84 mmol, 1.05eq) and K₂CO₃ (746.15 mg, 5.40 mmol, 2 eq) in DMSO (15 mL). Then themixture was stirred at 100° C. for 4 hours under N₂. TLC (Petroleumether: Ethyl acetate=3:1, Rf=0.5) showed no start material and a newspot. The residue was diluted with H₂O (50 mL) extracted with ethylacetate (60 mL×3). The combined organic layers were washed with brine 40mL, dried over anhydrous sodium sulfate, filtered and concentrated underreduced pressure to give a residue. The residue was purified by silicagel column chromatography (0 to 30% Ethyl acetate in Petroleum ether) togive tert-butyl4-[2-[4-(4-bromo-2-pyridyl)piperazin-1-yl]-1,1-difluoro-ethyl]piperidine-1-carboxylate(1 g, 1.94 mmol, 71.91% yield, 95% purity) as a white solid.

Exemplary Synthesis of Exemplary Compound 23:(7R)—N-{3-[2-(4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 23 was prepared in a manner analogous to ExemplaryCompound 21 using enantiomer 2 of4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid.

Step 1

To a solution of tert-butyl 4-methylenepiperidine-1-carboxylate (6.24 g,31.65 mmol, 1 eq) was added 9-BBN (0.5 M, 63.29 mL, 1 eq) at 25° C. Thereaction mixture was stirred at 80° C. for 1 hour under N₂. Aftercooling, 4-bromopyridine (5 g, 31.65 mmol, 1 eq), Pd(dppf)Cl2 (1.39 g,1.90 mmol, 0.06 eq), K2CO3 (6.56 g, 47.47 mmol, 1.5 eq), DMF (50 mL) andH2O (5 mL) were added to the reaction. The resultant mixture was heatedto 60° C. for 12 hours. LCMS showed desired MS. After cooling, thereaction mixture was diluted with water (100 mL) and extracted withethyl acetate (3×100 mL). The organic layer was washed with brine (2×100mL), dried over sodium sulfate and concentrated under reduced pressure.The residue was purified by silica gel column chromatography (0 to 40%ethyl acetate in petroleum ether) to afford tert-butyl4-(4-pyridylmethyl)piperidine-1-carboxylate (3.49 g, 11.66 mmol, 36.83%yield, 92.3% purity) as a pale yellow oil.

Step 2

To a solution of tert-butyl 4-(4-pyridylmethyl)piperidine-1-carboxylate(3.49 g, 12.63 mmol, 1 eq) in EtOH (50 mL) and HOAc (758.33 mg, 12.63mmol, 722.21 uL, 1 eq) was added PtO₂ (430.12 mg, 1.89 mmol, 0.15 eq) at25° C. Then the mixture was stirred at 70° C. for 24 hours under H₂ (50psi). TLC (PE:EA=1:1) showed starting material consumed and a new spotformed. After cooling, the reaction was filtered and filtrate wasconcentrated under reduced pressure to afford tert-butyl4-(4-piperidylmethyl)piperidine-1-carboxylate (3.9 g, crude) as a brownoil.

Step 3

To a solution of tert-butyl4-(4-piperidylmethyl)piperidine-1-carboxylate (3.7 g, 13.10 mmol, 1.21eq) and 2-(2,6-dioxo-3-piperidyl)-5-fluoro-isoindoline-1,3-dione (3 g,10.86 mmol, 1 eq) in DMSO (40 mL) was added DIEA (5.61 g, 43.44 mmol,7.57 mL, 4 eq). After addition, the reaction mixture was stirred at 100°C. for 2 hours. TLC (petroleum ether: ethyl acetate=1:1) showed a newspot. After cooling, the reaction was diluted with ethyl acetate (200mL) and washed with brine (3×100 mL). The organic layer was dried oversodium sulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (0 to 50% ethyl acetate inpetroleum ether) to afford tert-butyl4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]piperidine-1-carboxylate(2.86 g, 4.41 mmol, 40.58% yield, 83% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 24:(7R)—N-[3-(2-{4-[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)azetidin-3-yl]piperazin-1-yl}pyridin-4-yl)-1H-indazol-5-yl]-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of tert-butyl 3-oxoazetidine-1-carboxylate (170 mg, 993.03umol, 1 eq) and 5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole(305.97 mg, 943.38 umol, 0.95 eq) in HOAc (1 mL) and MeOH (10 mL) wasstirred at 20° C. for 20 minutes, then was added borane;2-methylpyridine (212.43 mg, 1.99 mmol, 2 eq). Then the mixture wasstirred at 30° C. for 16 hours under N₂. TLC(Dichloromethane:Methanol=10:1, Rf=0.3) showed no start material and anew spot. The residue was concentrated under reduced pressure to give aresidue. The residue was purified by silica gel column chromatography (0to 20% Dichloromethane in Methanol) to give tert-butyl3-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]azetidine-1-carboxylate(460 mg, 774.13 umol, 77.96% yield, 80.7% purity) as a yellow solid.

Step 2

To a solution of tert-butyl3-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]azetidine-1-carboxylate(460 mg, 959.27 umol, 1 eq) in DCM (3 mL) was added TFA (4.25 g, 37.28mmol, 2.76 mL, 38.86 eq) and then stirred at 20° C. for 1 hr. TLC(Dichloromethane:Methanol=5:1, Rf=0.01) showed no start material and anew spot. The residue was concentrated under reduced pressure to give3-[2-[4-(azetidin-3-yl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(360 mg, crude, TFA) as a yellow solid.

Step 3

To a solution of1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carbaldehyde(292.06 mg, 790.69 umol, 1 eq) and3-(2-(4-(azetidin-3-yl)piperazin-1-yl)pyridin-4-yl)-5-nitro-1H-indazole(300.00 mg, 790.69 umol, 1 eq) in HOAc (1 mL) and MeOH (20 mL) wasstirred at 20° C. for 20 minutes, then was added borane;2-methylpyridine (169.15 mg, 1.58 mmol, 2 eq). Then the mixture wasstirred at 30° C. for 16 hours under N₂. TLC(Dichloromethane:Methanol=5:1, Rf=0.1) showed no start material and anew spot. The residue was concentrated under reduced pressure to give aresidue. The residue was purified by silica gel column chromatography (0to 50% Dichloromethane in Methanol) to give2-(2,6-dioxopiperidin-3-yl)-5-(4-((3-(4-(4-(5-nitro-1H-indazol-3-yl)pyridin-2-yl)piperazin-1-yl)azetidin-1-yl)methyl)piperidin-1-yl)isoindoline-1,3-dione(360 mg, 412.67 umol, 52.19% yield, 84% purity) as a yellow solid.

Step 4

To a solution of2-(2,6-dioxo-3-piperidyl)-5-[4-[[3-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]azetidin-1-yl]methyl]-1-piperidyl]isoindoline-1,3-dione(360 mg, 491.28 umol, 1 eq) in EtOH (10 mL) and H₂O (5 mL) was added Fe(137.18 mg, 2.46 mmol, 5 eq), NH₄Cl (131.39 mg, 2.46 mmol, 5 eq). Thenthe mixture was stirred at 90° C. for 1 hour under N₂. LCMS showeddesired product. The residue was concentrated under reduced pressure togive a residue. The residue was purified by prep-HPLC (PhenomenexGemini-NX 150*30 mm*5 um: water (0.05% HCl)-ACN; B %: 10%-40%, 10 min)to afford5-[4-[[3-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]azetidin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(177 mg, 231.70 umol, 47.16% yield, 92% purity) as a yellow solid.

Step 5

To a solution of5-[4-[[3-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]azetidin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(78 mg, 110.98 umol, 1 eq) and(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(23.22 mg, 110.98 umol, 1 eq) in DMF (4 mL) was stirred at 20° C. for 10minutes, then was added DIEA (71.72 mg, 554.92 umol, 96.65 uL, 5 eq) andHATU (42.20 mg, 110.98 umol, 1 eq). Then the mixture was stirred at 30°C. for 16 hours under N₂. LCMS showed desired product. The residue wasconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 um;mobile phase: [water (0.225% FA)-ACN]; B %: 0%-30%, 35 min) to give(7R)—N-[3-[2-[4-[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]azetidin-3-yl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(19.1 mg, 21.21 umol, 19.11% yield, 99.27% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 25:(7R)—N-{3-[2-(4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of tert-butyl 4-(2-chloroethyl)piperazine-1-carboxylate(200 mg, 804.02 umol, 1 eq) in DCM (2 mL) added TFA (3.08 g, 27.01 mmol,2 mL, 33.60 eq) and then the mixture was stirred at 20° C. for 1 hour.TLC (Dichloromethane:Methanol=10:1, Rf=0.02) showed no start materialand a new spot. The residue was concentrated under reduced pressure togive 1-(2-chloroethyl)piperazine (119 mg, crude, TFA) as a Colorlessoil.

Step 2

To a solution of 1-(2-chloroethyl)piperazine (119 mg, 800.63 umol, 1 eq)and1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperidine-4-carbaldehyde(295.73 mg, 800.63 umol, 1 eq) in HOAC (1 mL) and MeOH (10 mL) wasstirred at 20° C. for 20 min, then was added borane; 2-methylpyridine(171.27 mg, 1.60 mmol, 2 eq). Then the mixture was stirred at 30° C. for16 hours under N₂. TLC (Dichloromethane:Methanol=10:1, Rf=0.3) showed nostart material and a new spot. The residue was concentrated underreduced pressure to give a residue. The residue was purified by silicagel column chromatography (0 to 25% Dichloromethane in Methanol) to give5-[4-[[4-(2-chloroethyl)piperazin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(390 mg, 769.12 umol, 96.06% yield, 99% purity) as a yellow solid.

Step 3

To a solution of(7R)-4,5,7-trimethyl-N-[3-(2-piperazin-1-yl-4-pyridyl)-1H-indazol-5-yl]-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(40 mg, 82.38 umol, 1 eq) and5-[4-[[4-(2-chloroethyl)piperazin-1-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(45 mg, 89.64 umol, 1.09 eq) and DIEA (106.47 mg, 823.82 umol, 143.49uL, 10 eq) and KI (136.75 mg, 823.82 umol, 10 eq) in MeCN (10 mL). Thenthe mixture was stirred at 80° C. for 4 hours under N₂. LCMS showeddesired product. The residue was diluted with H₂O (20 mL) and extractedwith ethyl acetate (20 mL×3). The combined organic layers were washedwith brine (15 mL), dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 um;mobile phase: [water (0.225% LA)-ACN]; B %: 0%-30%, 35 min) to afford(7R)—N-[3-[2-[4-[2-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]piperazin-1-yl]ethyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(8.8 mg, 9.04 umol, 10.97% yield, 97.66% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 26:(7R)—N-(3-{2-[6-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)-2,6-diazaspiro[3.3]heptan-2-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a mixture of 4-bromo-2-fluoro-pyridine (1 g, 5.68 mmol, 1 eq) andtert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate; oxalic acid (1.38g, 2.84 mmol, 0.5 eq) in DMSO (10 mL) was added K₂CO₃ (2.36 g, 17.04mmol, 70.14 mL, 3 eq) in one portion at 100° C. under N₂. The mixturewas stirred at 100° C. for 2 hours to give yellow solution. TLC and LCMSshowed the reaction was completed. The mixture was cooled to 20° C. andconcentrated in reduced pressure at 20° C. The residue was poured intowater (10 mL). The aqueous phase was extracted with ethyl acetate (10mL*4). The combined organic phase was washed with brine (10 mL*2), driedwith anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residuewas purified by silica gel chromatography (Petroleum ether: Ethylacetate=10:1, Rf=0.56, 12 g, 0-50% (10 min) of Ethyl acetate inPetroleum ether, 50% (5 min) of Ethyl acetate in Petroleum ether) togive tert-butyl6-(4-bromo-2-pyridyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (1.3 g,3.67 mmol, 64.61% yield) as a white solid.

Step 2

To a mixture of2-[(3-bromo-5-nitro-indazol-1-yl)methoxy]ethyl-trimethyl-silane (2 g,5.37 mmol, 1 eq),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(2.05 g, 8.06 mmol, 1.5 eq) and Pd(dppf)Cl₂ (196.54 mg, 268.61 umol,0.05 eq) in dioxane (20 mL) was added KOAc (1.58 g, 16.12 mmol, 3 eq) inone portion at 25° C. under N₂. The mixture was stirred at 100° C. for 1hour. TLC showed the reaction was completed. The mixture was cooled to25° C., filtered and concentrated in vacuum to give[5-nitro-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]boronic acid (1.8g, crude) as a black brown solid. The crude product was used into thenext step without purification.

Step 3

To a mixture of tert-butyl6-(4-bromo-2-pyridyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (1.3 g,3.67 mmol, 1 eq),[5-nitro-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]boronic acid (1.73g, 5.14 mmol, 1.4 eq) and 4-ditert-butylphosphanyl-N,N-dimethyl-aniline;dichloropalladium (259.85 mg, 366.98 umol, 259.85 uL, 0.1 eq) in EtOH(10 mL) and H₂O (4 mL) was added KOAc (1.08 g, 11.01 mmol, 3 eq) in oneportion at 25° C. under N₂. The mixture was stirred at 100° C. for 2hours. LCMS showed the reaction was completed. The mixture was cooled to20° C. and concentrated in reduced pressure. The residue was poured intowater (10 mL). The aqueous phase was extracted with ethyl acetate (10mL*3). The combined organic phase was washed with brine (10 mL*2), driedwith anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residuewas purified by silica gel chromatography (20 g, 0-40% (10 min) of Ethylacetate in Petroleum ether, 40% (10 min) of Ethyl acetate in Petroleumether) to give tert-butyl6-[4-[5-nitro-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]-2-pyridyl]-2,6-diazaspiro[3.3]heptane-2-carboxylate(1.2 g, 2.12 mmol, 57.70% yield) as a yellow solid.

Step 4

To a mixture of tert-butyl6-[4-[5-nitro-1-(2-trimethylsilylethoxymethyl)indazol-3-yl]-2-pyridyl]-2,6-diazaspiro[3.3]heptane-2-carboxylate(600 mg, 1.06 mmol, 1 eq) in DCM (5 mL) was added TFA (362.16 mg, 3.18mmol, 235.17 uL, 3 eq) in one portion at 20° C. The mixture was stirredat 20° C. for 2 hours. The mixture added dioxane (10 mL) and NH₃.H₂O(556.55 mg, 6.35 mmol, 611.60 uL, 40% purity, 6 eq) in one portion at20° C., The mixture was stirred at 20° C. for 2 hours. LCMS showed thereaction was completed. The residue was poured into water (5 mL), theaqueous phase was extracted with DCM (5 mL*3). The combined organicphase was washed with brine (5 mL*2), dried with anhydrous Na₂SO₄,filtered and concentrated in vacuum to give3-[2-(2,6-diazaspiro[3.3]heptan-2-yl)-4-pyridyl]-5-nitro-1H-indazole(300 mg, 561.92 umol, 53.08% yield, 63% purity) as a yellow solid.

Step 5

To a mixture of3-[2-(2,6-diazaspiro[3.3]heptan-2-yl)-4-pyridyl]-5-nitro-1H-indazole(300 mg, 891.94 umol, 1 eq) and1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperidine-4-carbaldehyde(329.45 mg, 891.94 umol, 1 eq) in MeOH (10 mL) was added CH₃COOH (53.56mg, 891.94 umol, 51.01 uL, 1 eq) and borane; 2-methylpyridine (190.80mg, 1.78 mmol, 2 eq) in one portion at 20° C. under N₂. The mixture wasstirred at 30° C. for 16 hours. LCMS showed there was desired MS. Theresidue was poured into water (10 mL). The aqueous phase was extractedwith ethyl acetate (10 mL*3). The combined organic phase was washed withbrine (10 mL*2), dried with anhydrous Na₂SO₄, filtered and concentratedin vacuum. The residue was purified by silica gel chromatography(Dichloromethane:Methanol=10:1, Rf=0.32, 0-10% (10 min) of Methanol inDichloromethane, 10% (5 min) of Methanol in Dichloromethane) to give2-(2,6-dioxo-3-piperidyl)-5-[4-[[2-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]-2,6-diazaspiro[3.3]heptan-6-yl]methyl]-1-piperidyl]isoindoline-1,3-dione(600 mg, 565.45 umol, 63.40% yield, 65% purity) as a yellow gum.

Step 6

To a mixture of2-(2,6-dioxo-3-piperidyl)-5-[4-[[2-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]-2,6-diazaspiro[3.3]heptan-6-yl]methyl]-1-piperidyl]isoindoline-1,3-dione(600 mg, 869.92 umol, 1 eq) and Fe (242.90 mg, 4.35 mmol, 5 eq) in EtOH(10 mL), H₂O (5 mL) and DCM (5 mL) was added NH₄Cl (232.67 mg, 4.35mmol, 5 eq) in one portion at 20° C. under N₂. The mixture was stirredat 80° C. for 30 minutes. LCMS showed the reaction was completed. Theresidue was filtered with ethyl acetate (10 mL*5), and solution wasconcentrated in vacuum. The crude product was purified by reversed-phaseHPLC (Column: YMC-Triart Prep C18 150*40 mm*7 um; Condition: water(0.05% HCl)-ACN; Begin B: 5; End B: 35; FlowRate: 60 mL/min; GradientTime: 25 min; 100% B Hold Time: 2 min) to give5-(4-((6-(4-(5-amino-1H-indazol-3-yl)pyridin-2-yl)-2,6-diazaspiro[3.3]heptan-2-yl)methyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione(110 mg, 166.07 umol, 19.09% yield, 99.6% purity) as a yellow solid.

Step 7

To a solution of(7R)-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxylic acid(34.88 mg, 166.73 umol, 1 eq) and5-[4-[[2-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]-2,6-diazaspiro[3.3]heptan-6-yl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(110 mg, 166.73 umol, 1 eq) in DMF (2 mL) was stirred at 0° C. for 10minutes, then was added DIPEA (107.74 mg, 833.67 umol, 145.21 uL, 5 eq)and HATU (63.40 mg, 166.73 umol, 1 eq), Then the mixture was stirred at25° C. for 16 hours under N₂. LCMS showed desired MS. The residue waspoured into water (2 mL). The aqueous phase was extracted with ethylacetate (2 mL*3). The combined organic phase was washed with brine (2mL*2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum.The crude product was purified by reversed-phase HPLC (Column:3_Phenomenex Luna C18 75*30 mm*3 um; Condition: water (0.225% FA)-ACN;Begin B: 0; End B: 30; FlowRate: 25 mL/min; Gradient Time: 35 minutes;100% B Hold Time: 3 minutes) to give(7R)—N-[3-[2-[6-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-2,6-diazaspiro[3.3]heptan-2-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7-trimethyl-7H-tetrazolo[1,5-a]pyrimidine-6-carboxamide(29 mg, 34.01 umol, 20.40% yield, 99.78% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 27:(7S)—N-{3-[2-(4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamideuors

Step 1

To a solution of2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-yl-isoindoline-1,3-dione (2 g,4.38 mmol, 1 eq, TFA) and tert-butyl 4-formylpiperidine-1-carboxylate(1.40 g, 6.57 mmol, 1.5 eq) in MeOH (20 mL) was added NaOAc (1.08 g,13.15 mmol, 3 eq), HOAc (131.59 mg, 2.19 mmol, 125.32 uL, 0.5 eq) andNaBH₃CN (826.20 mg, 13.15 mmol, 3 eq) in one portion at 20° C. under N₂.The solution was stirred at 20° C. for 1 hour. TLC (DCM:MeOH=10:1,Rf=0.43) showed the reaction was completed, and a main spot (Rf=0.43)was showed on TLC. The residue was poured into water (20 mL). Theaqueous phase was extracted with ethyl acetate (3×20 mL). The combinedorganic phase was washed with brine (2×20 mL), dried with anhydrousNa₂SO₄, filtered and concentrated in vacuum. The residue was purified bysilica gel chromatography (column: 40 g, 100-200 mesh silica gel, 0-50%(5 min) of Ethyl acetate in Petroleum ether, 50-100% (10 min) of Ethylacetate in Petroleum ether) to give tert-butyl4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]methyl]piperidine-1-carboxylate(2.2 g, 3.99 mmol, 90.98% yield, 97.8% purity) as a yellow gum.

Step 2

To a solution of tert-butyl4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]methyl]piperidine-1-carboxylate(2.2 g, 4.08 mmol, 1 eq) in DCM (2 mL) was added TFA (1.39 g, 12.23mmol, 905.58 uL, 3 eq) in one portion at 20° C. under N₂. The mixturewas stirred at 20° C. for 30 minutes to give yellow solution. TLC(DCM:MeOH=10:1, Rf=0.43) showed the starting material was completed. Thesolution was concentrated in vacuum to give2-(2,6-dioxo-3-piperidyl)-5-[4-(4-piperidylmethyl)piperazin-1-yl]isoindoline-1,3-dione(3 g, 3.73 mmol, 91.61% yield, 97.3% purity, 3TFA) as a yellow gum.

Step 3

To a mixture of 5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole (500mg, 1.54 mmol, 1 eq) and 2-chloroacetaldehyde (520 mg, 2.65 mmol, 426.23uL, 40% purity, 1.72 eq) in DCE (20 mL) and MeOH (20 mL) was addedCH₃COOH (0.1 mL) and NaBH₃CN (290.63 mg, 4.62 mmol, 3 eq) in one portionat 20° C. under N₂. The mixture was stirred at 20° C. for 2 hours. LCMSshowed there was desired MS. The residue was poured into water (10 mL).The aqueous phase was extracted with ethyl acetate (10 mL*3). Thecombined organic phase was washed with brine (10 mL*2), dried withanhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue waspurified by silica gel chromatography (Dichloromethane:Methanol=10:1,Rf=0.27, 0-50% (15 min) of Ethyl acetate in Petroleum ether, 10% (5 min)of Ethyl acetate in Petroleum ether) to give3-[2-[4-(2-chloroethyl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(350 mg, 904.78 umol, 58.69% yield) as a yellow solid.

Step 4

To a mixture of3-[2-[4-(2-chloroethyl)piperazin-1-yl]-4-pyridyl]-5-nitro-1H-indazole(350 mg, 904.78 umol, 1 eq) and2-(2,6-dioxo-3-piperidyl)-5-[4-(4-piperidylmethyl)piperazin-1-yl]isoindoline-1,3-dione(397.66 mg, 508.79 umol, 5.62e-1 eq, 3TFA) in MeCN (10 mL) was added KI(300.39 mg, 1.81 mmol, 2 eq) and DIPEA (233.87 mg, 1.81 mmol, 315.19 uL,2 eq) in one portion at 20° C. under N₂. The mixture was stirred at 80°C. for 16 hours. LCMS showed there was no starting material. The residuewas added H₂O (10 mL) and the solid formed was filtered under vacuum togive2-(2,6-dioxo-3-piperidyl)-5-[4-[[1-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-4-piperidyl]methyl]piperazin-1-yl]isoindoline-1,3-dione(600 mg, crude) as a yellow solid.

Step 5

To a mixture of2-(2,6-dioxo-3-piperidyl)-5-[4-[[1-[2-[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-4-piperidyl]methyl]piperazin-1-yl]isoindoline-1,3-dione(600 mg, 759.61 umol, 1 eq) and NH₄Cl (203.16 mg, 3.80 mmol, 5 eq) inEtOH (10 mL), H₂O (2 mL) and DCM (2 mL) was added Fe (212.10 mg, 3.80mmol, 5 eq) in one portion at 20° C. under N₂. The mixture was stirredat 80° C. for 1 hour. LCMS showed the reaction was completed. Theresidue was filtered with ethyl acetate (10 mL*5) and solution wasconcentrated in vacuum. The crude product was purified by reversed-phaseHPLC (Column: 3_Phenomenex Luna C18 75*30 mm*3 um; Condition: water(0.05% HCl)-ACN; Begin B: 10.End B: 80; FlowRate: 25 mL/min; GradientTime: 35 min; 100% B Hold Time: 1 minute) to give5-[4-[[1-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-4-piperidyl]methyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(180 mg, 232.14 umol, 30.56% yield, 98% purity) as a yellow solid.

Step 6

To a mixture of5-[4-[[1-[2-[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]ethyl]-4-piperidyl]methyl]piperazin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(90 mg, 118.44 umol, 1 eq) and4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylic acid (43.11mg, 177.66 umol, 92% purity, 1.5 eq) in DMF (5 mL) was added DIEA (45.92mg, 355.31 umol, 61.89 uL, 3 eq) in one portion and HATU (45.03 mg,118.44 umol, 1 eq) at 25° C. under N₂. The mixture was stirred at 25° C.for 16 hours to give yellow solution. LCMS showed desired product. Theresidue was concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30mm*3 um; mobile phase: [water (0.1M FANH4)-ACN]; B %: 0%-30%, 40 min) toaffordN-[3-[2-[4-[2-[4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazin-1-yl]methyl]-1-piperidyl]ethyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7,7-tetramethyl-tetrazolo[1,5-a]pyrimidine-6-carboxamide(18.9 mg, 19.17 umol, 16.18% yield, 97.88% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 28:(7S)—N-(3-{2-[(3S)-4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 28 was prepared in a manner analogous to ExemplaryCompound 27.

Exemplary Synthesis of Exemplary Compound 29:N-{3-[2-(4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Step 1

To a solution of 5-nitro-3-(2-piperazin-1-yl-4-pyridyl)-1H-indazole (500mg, 1.54 mmol, 1 eq) and tert-butyl 4-formylpiperidine-1-carboxylate(450 mg, 2.11 mmol, 1.37 eq) in HOAc (1 mL) and MeOH (10 mL) was addedborane; 2-methylpyridine (329.78 mg, 3.08 mmol, 2 eq). After addition,the reaction solution was stirred at 20° C. for 2 hours. LCMS showeddesired MS. The reaction was diluted with brine (100 mL). The solid wascollected by filtration and dried under reduced pressure to affordtert-butyl4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(730 mg, 1.20 mmol, 78.07% yield, 86% purity) as a yellow solid.

Step 2

To a solution of tert-butyl4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]piperidine-1-carboxylate(730 mg, 1.40 mmol, 1 eq) in DCM (10 mL) was added TFA (7.70 g, 67.53mmol, 5 mL, 48.25 eq). After addition, the reaction solution was stirredat 20° C. for 3 hours. LCMS showed starting material consumed anddesired MS was detected. The reaction was concentrated under reducedpressure to afford5-nitro-3-[2-[4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(589 mg, crude) as a yellow gum. The crude product was used for nextstep directly.

Step 3

To a solution of5-nitro-3-[2-[4-(4-piperidylmethyl)piperazin-1-yl]-4-pyridyl]-1H-indazole(589 mg, 1.40 mmol, 1 eq) and1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperidine-4-carbaldehyde(619.39 mg, 1.68 mmol, 1.2 eq) in HOAc (5 mL) and MeOH (50 mL) was addedborane; 2-methylpyridine (298.94 mg, 2.79 mmol, 2 eq). After addition,the reaction mixture was stirred at 20° C. for 12 hours. LCMS showedstarting material consumed and desired MS detected. The reaction mixturewas dilute with water (50 mL) and extracted with 10% methanol indichloromethane (3×50 mL). The organic layer was dried over sodiumsulfate and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (0 to 40% methanol indichloromethane) to afford2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]isoindoline-1,3-dione(1 g, 1.25 mmol, 89.35% yield, 96.75% purity) as a yellow solid.

Step 4

To a solution of2-(2,6-dioxo-3-piperidyl)-5-[4-[[4-[[4-[4-(5-nitro-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]isoindoline-1,3-dione(500 mg, 645.27 umol, 1 eq) in EtOH (5 mL) and H₂O (2 mL) was added Le(180.18 mg, 3.23 mmol, 5 eq) and NH4Cl (172.58 mg, 3.23 mmol, 5 eq).After addition, the reaction mixture was stirred at 80° C. for 1 hour.LCMS showed desired MS. After cooling, the reaction mixture was filteredand filtrate was concentrated under reduced pressure. The residue wastriturated with 10% methanol in dichloromethane (20 mL) and filtered.The filtrate was concentrated under reduced pressure to afford5-[4-[[4-[[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(110 mg, 143.24 umol, 22.20% yield, 97% purity) as a yellow solid.

Step 5

To a mixture of5-[4-[[4-[[4-[4-(5-amino-1H-indazol-3-yl)-2-pyridyl]piperazin-1-yl]methyl]-1-piperidyl]methyl]-1-piperidyl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione(100 mg, 134.25 umol, 1 eq) and4,5,7,7-tetramethyltetrazolo[1,5-a]pyrimidine-6-carboxylic acid (56.19mg, 201.37 umol, 80% purity, 1.5 eq) in DMF (5 mL) was added DIEA (52.05mg, 402.75 umol, 70.15 uL, 3 eq) in one portion and HATU (51.05 mg,134.25 umol, 1 eq) at 25° C. under N₂. The mixture was stirred at 25° C.for 16 hours to give yellow solution. LCMS showed desired product. Theresidue was concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: 3_Phenomenex Luna C18 75*30mm*3 um; mobile phase: [water (0.1M FANH4)-ACN]; B %: 0%-30%, 40 min) toaffordN-[3-[2-[4-[[1-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]-4-piperidyl]methyl]-4-piperidyl]methyl]piperazin-1-yl]-4-pyridyl]-1H-indazol-5-yl]-4,5,7,7-tetramethyl-tetrazolo[1,5-a]pyrimidine-6-carboxamide(12 mg, 12.25 umol, 9.13% yield, 97% purity) as a yellow solid.

Exemplary Synthesis of Exemplary Compound 30:N-(3-{2-[(3S)-4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 30 was prepared in a manner analogous to ExemplaryCompound 29.

Exemplary Synthesis of Exemplary Compound 31:(7S)—N-(3-{2-[(3S)-4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 30 was prepared in a manner analogous to ExemplaryCompound 29.

Exemplary Synthesis of Exemplary Compound 32:N-(3-{2-[(2S)-2-{[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]methyl}morpholin-4-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 30 was prepared in a manner analogous to ExemplaryCompound 29.

Step 1

To a mixture of tert-butyl(2R)-2-(hydroxymethyl)morpholine-4-carboxylate (2 g, 9.21 mmol, 1 eq) inDCM (20 mL) was added TFA (4.62 g, 40.52 mmol, 3 mL, 4.40 eq) in oneportion at 20° C. under N₂. The mixture was stirred at 20° C. for 30minutes. TLC showed the reaction was completed. The solution wasconcentrated in vacuum to give [(2R)-morpholin-2-yl]methanol (2 g, 8.65mmol, 93.98% yield, TFA) as a colourless oil.

Step 2

To a mixture of 4-bromo-2-fluoro-pyridine (1.52 g, 8.65 mmol, 1 eq) and[(2R)-morpholin-2-yl]methanol (2 g, 8.65 mmol, 1 eq, TFA) in DMSO (10mL) was added K₂CO₃ (11.96 g, 86.52 mmol, 70.14 mL, 10 eq) in oneportion at 100° C. under N₂. The mixture was stirred at 100° C. for 2hours to give yellow solution. TLC (Petroleum ether: Ethyl acetate=1:1,Rf=0.56) showed the reaction was completed. The mixture was cooled to20° C. and concentrated in reduced pressure at 20° C. The residue waspoured into water (50 mL). The aqueous phase was extracted with ethylacetate (50 mL*4). The combined organic phase was washed with brine (50mL*2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum.The residue was purified by silica gel chromatography (Petroleum ether:Ethyl acetate=1:1, Rf=0.56, 20 g, 0-50% (10 min) of Ethyl acetate inPetroleum ether, 50% (10 min) of Ethyl acetate in Petroleum ether) togive [(2R)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methanol (1.5 g, 5.49mmol, 63.48% yield) as a yellow oil.

Step 3

To a mixture of [(2R)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methanol (1.5g, 5.49 mmol, 1 eq) in DCM (15 mL) was added TEA (1.11 g, 10.98 mmol,1.53 mL, 2 eq) and 4-methylbenzenesulfonyl chloride (1.57 g, 8.24 mmol,1.5 eq) in one portion at 0° C. under N₂. The mixture was stirred at 20°C. for 16 hours. LCMS showed the reaction was completed. The residue waspoured into water (10 mL). The aqueous phase was extracted with DCM (20mL*2). The combined organic phase was washed with brine (20 mL*2), driedwith anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residuewas purified by silica gel chromatography (12 g, 0-100% (20 min) ofEthyl acetate in Petroleum ether) to give[(2R)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methyl4-methylbenzenesulfonate (2.1 g, 4.91 mmol, 89.52% yield) as acolourless oil

Step 4

To a mixture of [(2R)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methyl4-methylbenzenesulfonate (2.1 g, 4.91 mmol, 1 eq) and tert-butylpiperazine-1-carboxylate (1.83 g, 9.83 mmol, 2 eq) in MeCN (20 mL) wasadded KI (1.63 g, 9.83 mmol, 2 eq) and DIPEA (1.27 g, 9.83 mmol, 1.71mL, 2 eq) in one portion at 20° C. under N₂. The mixture was stirred at100° C. for 2 hours. TLC showed the reaction was completed. The mixturewas cooled to 20° C. and concentrated in reduced pressure at 20° C. Theresidue was poured into water (20 mL). The aqueous phase was extractedwith ethyl acetate (20 mL*3). The combined organic phase was washed withbrine (20 mL*2), dried with anhydrous Na₂SO₄, filtered and concentratedin vacuum. The residue was purified by silica gel chromatography (12 g,0-20% (5 min) of Ethyl acetate in Petroleum ether, 20% (15 minutes) ofEthyl acetate in Petroleum ether) to give tert-butyl4-[[(2S)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methyl]piperazine-1-carboxylate(1.1 g, 2.22 mmol, 45.14% yield, 89% purity) as a yellow gum.

Step 5

To a mixture of tert-butyl4-[[(2S)-4-(4-bromo-2-pyridyl)morpholin-2-yl]methyl]piperazine-1-carboxylate(1.1 g, 2.22 mmol, 89% purity, 1 eq), Pin₂B₂ (1.13 g, 4.44 mmol, 2 eq)and KOAc (653.06 mg, 6.65 mmol, 3 eq) in dioxane (300 mL) was addedPd(dppf)Cl₂ (81.15 mg, 110.91 umol, 0.05 eq) in one portion at 25° C.under N₂. The mixture was stirred at 100° C. for 1 hour. TLC showed thereaction was completed. The mixture was cooled to 25° C., filtered andconcentrated in vacuum to give[2-[(2S)-2-[(4-tert-butoxycarbonylpiperazin-1-yl)methyl]morpholin-4-yl]-4-pyridyl]boronicacid (1 g, 1.75 mmol, 78.78% yield, 71% purity) as a black brown solid.

Exemplary Synthesis of Exemplary Compound 33:N-{3-[2-(4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 33 was prepared in a manner analogous to ExemplaryCompound 27.

Exemplary Synthesis of Exemplary Compound 34:N-(3-{2-[(3S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4-yl]methyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 34 was prepared in a manner analogous to ExemplaryCompound 1.

Exemplary Synthesis of Exemplary Compound 35:N-{3-[2-(4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 35 was prepared in a manner analogous to ExemplaryCompound 27.

Exemplary Synthesis of Exemplary Compound 36:N-(3-{2-[(3S)-4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide

Exemplary Compound 35 was prepared in a manner analogous to ExemplaryCompound 25.

Protein Level Control

This description also provides methods for the control of protein levelswithin a cell. The method is based on the use of compounds as describedherein such that degradation of the target protein LRRK2 in vivo willresult in the reducing the amount of the target protein in a biologicalsystem, preferably to provide a particular therapeutic benefit.

The following examples are used to assist in describing the presentdisclosure, but should not be seen as limiting the present disclosure inany way.

In certain embodiments, the description provides the following exemplaryLRRK2-degrading bifunctional molecules (compounds of Table 1 orCompounds 1-36), including salts, polymorphs, analogs, derivatives, anddeuterated forms thereof.

Assay for Testing LRRK2 Degradation Driven by Compounds as DescribedHerein,

The assay measures the degradation of wildtype and G2019S LRRK2 taggedwith a HiBit tag on the C-terminus of the protein that was expressedfrom a mammalian expression vector, driven by the ubiquitin promoter inHEK293 cells. Each compound dose-response was repeated on three separatedays, on three separate plates each day.

Plasmid Preparation. Transfection mixes were assembled as follows andincubated for 30 minutes at room temperature. In a 15 mL tube, 5.25 mLOpti-MEM (no additions) was mixed with 17 μL Firefly Luciferase plasmidat 1 μg/μL and 158 μL WT plasmid DNA at 1 μg/μL (175 μg total DNA) weremixed by flicking. In a new 15 mL tube, 5.25 mL OptiMEM was mixed with17 μL Firefly Luciferase plasmid at 1 μg/μL and 158 μL G2019S plasmidDNA at 1 μg/μL (175 μg total DNA) were mixed by flicking. X-tremeGene HPwas mixed thoroughly using a vortex. Next, 175 μL was added to each tubeand flicked to mix. Both tubes were left to incubate for 30 minutes atroom temperature.

While the transfection mixes were incubating, HEK293 cells acquired fromATCC (ATCC CRL-1573) were harvested with trypsin. Once cells aredetached, the cells were resuspended in 12 mL OptiMEM+5% FBS andtransferred to a 50 mL tube. The cells were mixed well and counted.Using OptiMEM+5% FBS, the cells were diluted in two 250 mL conical tubesat 0.71×10⁶ cells/mL in 70 mL. One tube was labeled “WT” and the other“G2019S”. The WT and G2019S transfection mixes were added dropwise tothe corresponding 250 mL tubes. The tubes were mixed first by pipettingthen by swirling. The tubes were incubated at room temperature for atleast 5 minutes.

Each tube was swirled before dispensing and after every three plates.Seventy microliters of cells were dispensed with WT or G2019S DNA toseven plates each. Three plates of each were tested with compound plateone (preparation described below) and three plates of each were testedwith compound plate two (preparation described below). The first platefrom each set served as a “prime” plate and was not used to testcompounds. Each plate was incubated in the hood for 10 minutes beforeplacing in the 37° C. incubator for 24 hours.

Preparation of Compound and Assay Plates. Two compound plates were madeusing 96 well polypropylene plates. Compounds were made up at 10 mM andwere diluted to 1 mM in 30 μL. Each dose response curve included a wellof DMSO, as a negative control and for normalization, and a well of 0.5μM of Exemplary Compound 4 as a positive control. In addition to seventest compounds, each plate also included a dose response of ExemplaryCompound 4. The compound plates were spun down along at 1200 rpm for 2minutes.

The two compound plates were then mixed and 2 μL was diluted inintermediate plates having 248 μL of Opti-Mem in each well. Next, 10 μLdiluted compounds from the intermediate plates were added to each testplate (three WT and three G2019S plates per compound plate for a totalof 12 assay plates). The plates were incubated for 24 hours at 37° C.

All assay plates and all Nano-Glo Dual-Luciferase Reporter Assay Systemcomponents (except for the DLR substrate) were equilibrated to roomtemperature. Next, the luciferase buffer was mixed with the lyophilizedamber bottle until fully dissolved, and 75 μL of the luciferase mixturewas added to each well of each assay plate. The assay plates wereincubated for 10 minutes at room temperature with shaking for at least 5minutes, and then read on a plate reader.

Developing Plates and Analyzing Data. One milliliter of DLR substrateand 1 mL LgBiT Protein were added to the Stop and Glo buffer, and 75 μLof the mixture was added to each well of each plate. Optically clearseals were added to each plate and each plate was incubated for 20minutes with shaking for at least 10 minutes, and then read on a platereader.

As mentioned above, plates were fun in triplicate and assay repeatedtwice (total of 6 replicates per exemplary compound. Each cell wasexamined for firefly luciferase for cell number and viability andNanoluc for the LRRK2-HiBit quantification.

Ratio of (HiBit/luciferase)*1000 was determined and the datawas_normalized to % of DMSO median value. Curve fitting was performed oneach individual plate. The data for some of exemplary compounds of Table1 below is shown below in Table 2 in the columns labeled *G2019S DC50(nM) and **G2019S Dmax (%).

Exemplary Assay for Testing LRRK2 Degradation Driven by ExemplaryHetero-Bifunctional Compounds Designed to Target LRRK2

The assay measures the degradation of wildtype and G2019S LRRK2 taggedwith a HiBit tag on the C-terminus of the protein that was expressedfrom a mammalian expression vector, driven by the ubiquitin promoter inHEK293 cells. Each compound dose-response was repeated on two separatedays, on three separate plates each day.

Plasmid Preparation. Transfection mixes were assembled as follows andincubated for 30 minutes at room temperature. In a 15 mL tube, 5.25 mLOpti-MEM (no additions) was mixed with 17 μL Firefly Luciferase plasmidat 1 μg/μL and 158 μL WT plasmid DNA at 1 μg/μL (175 μg total DNA) weremixed by flicking. In a new 15 mL tube, 5.25 mL OptiMEM was mixed with17 μL Firefly Luciferase plasmid at 1 μg/μL and 158 μL G2019S plasmidDNA at 1 μg/μL (175 μg total DNA) were mixed by flicking. X-tremeGene HPwas mixed thoroughly using a vortex. Next, 175 μL was added to each tubeand flicked to mix. Both tubes were left to incubate for 30 minutes atroom temperature.

While the transfection mixes were incubating, HEK293 cells (acquiredfrom ATCC; ATCC CRL-1573) were harvested with trypsin. Once cells aredetached, the cells were resuspended in 12 mL OptiMEM+5% FBS andtransferred to a 50 mL tube. The cells were mixed well and counted.Using OptiMEM+5% FBS, the cells were diluted in two 250 mL conical tubesat 0.71×10⁶ cells/mL in 70 mL. One tube was labeled “WT” and the other“G2019S”. The WT and G2019S transfection mixes were added dropwise tothe corresponding 250 mL tubes. The tubes were mixed first by pipettingthen by swirling. The tubes were incubated at room temperature for atleast 5 minutes.

Each tube was swirled before dispensing and after every three plates.Seventy microliters of cells were dispensed with WT or G2019S DNA toseven plates each. Three plates of each were tested with compound plateone (preparation described below) and three plates of each were testedwith compound plate two (preparation described below). The first platefrom each set served as a “prime” plate and was not used to testcompounds. Each plate was incubated in the hood for 10 minutes beforeplacing in the 37° C. incubator for 24 hours.

Preparation of Compound and Assay Plates. Two compound plates were madeusing 96 well polypropylene plates. Compounds were made up at 10 mM andwere diluted to 1 mM in 30 μL. Each dose response curve included a wellof DMSO, as a negative control and for normalization, and a well of 0.5μM of Exemplary Compound 4 as a positive control. In addition to seventest compounds, each plate also included a dose response of ExemplaryCompound 4. The compound plates were spun down along at 1200 rpm for 2minutes.

The two compound plates were then mixed and 2 μL was diluted inintermediate plates having 248 μL of Opti-Mem in each well. Next, 10 μLdiluted compounds from the intermediate plates were added to each testplate (three WT and three G2019S plates per compound plate for a totalof 12 assay plates). The plates were incubated for 24 hours at 37° C.

All assay plates and all Nano-Glo Dual-Luciferase Reporter Assay Systemcomponents (except for the DLR substrate) were equilibrated to roomtemperature. Next, the luciferase buffer was mixed with the lyophilizedamber bottle until fully dissolved, and 75 μL of the luciferase mixturewas added to each well of each assay plate. The assay plates wereincubated for 10 minutes at room temperature with shaking for at least 5minutes, and then read on a plate reader.

Developing Plates and Analyzing Data. One milliliter of DLR substrateand 1 mL LgBiT Protein were added to the Stop and Glo buffer, and 75 μLof the mixture was added to each well of each plate. Optically clearseals were added to each plate and each plate was incubated for 20minutes with shaking for at least 10 minutes, and then read on a platereader.

As mentioned above, plates were run in triplicate and assay repeatedtwice (total of 6 replicates per exemplary compound. Each cell wasexamined for firefly luciferase for cell number and viability andNanoluc for the LRRK2-HiBit quantification.

Ratio of (HiBit/luciferase)*1000 was determined and the datawas_normalized to % of DMSO median value. Curve fitting was performed oneach individual plate. The data for exemplary compounds of Table 1 belowis shown below in Table 2 in the G2019S DC50, G2019S Dmax, WT DC50 andWT Dmax columns.

Exemplary Assay for Testing LRRK2 Degradation Driven by ExemplaryHetero-Bifunctional Compounds Designed to Target LRRK2

The assay measures the degradation of LRRK2 in cells where theC-terminus (3′) of the endogenous gene has been tagged with a HiBitsequence in HEK293 cells. The cells also express firefly luciferase,expressed from a Cytomegalovirus promoter and introduced into the HiBittagged cells and stably expressed. The Nano-Glo® Dual LuciferaseReporter Assay System (Promega™, Madison, Wis.) was utilized.

Day 1—Preparation of Compound and Assay Plates. Two sets of plates wereprepared: a triplicate set for the HiBit assay in white 384-well platesand a triplicate set of plate in black 384-well plates for the AlamarBlue cell viability assay. Briefly, the growth media (DMEM+Glutamax-10%fetal bovine serum-1% Penicillin-Streptomicin) from two T128 flasks wasaspirated from the flasks. Cells were washed with Dulbecco's PhosphateBuffered Saline (dPBS) and aspirated. Trypsin (3 mL per flask) was addedand the flasks were incubated for 2-3 minutes.

Ten mL of OptiMEM-10% fetal bovine-1% penicillin-streptomycin(hereinafter, “OptiMEM media”) was added to the flask and the cells andtransferred to a 50 mL conical tube. A cell count (25 ul of cell intoEffendorf vial +25 ul of Trypan Blue Stain) was performed and the celldensity adjusted to 15,000 cell/45 μl/well (3.33×10{circumflex over( )}5/mL) in OptiMEM media.

Forty-five microliters of the cell suspension (15,000 cells) wasaliquoted to each well of the white 384-well plate. The plates incubatedat room temperature for 10 minutes before being placed in the 37° C.+5%CO₂ incubator overnight

Day 2—Compound Treatment. Exemplary compounds were prepared at a 1 mMstarting concentration and 1:3 serial dilution for 11 points CRCprepared and stored in the freezer. The Master Compound Plate was thawedovernight at room temperature. DMSO (20 μL) was added into column 24 ofthe Master Compound Plate for negative control and 20 μL of 300 μM ofExemplary Compound 4 in column 23 as positive control.

Intermediate Compound Plate with 4% DMSO in OptiMEM Media. DMSO wasadded to warm OptiMEM media to achieve a 4% DMSO solution (approximately50 mL/plate). One-hundred microliters of the OptiMEM-4% DMSO wasaliquoted to each well of 384-Well Deep Well Microplates.

The Master Compound Plate and the Intermediate Compound Plate were spundown.

One microliter of compound from the Master Compound Plate wastransferred into the Intermediate plate (a 1:100 dilution). The dilutedmixture was mixed and 5 μL transferred into the assay plate (a 1:10dilution) for the final starting concentration of 1 μM. The TreatedAssay plates were incubated for 24 hours at 37° C.+5% CO₂. The MasterCompound Plate was sealed and store at room temperature for a second runthat was performed within a week.

Day 3—HiBit Assay. Five microliters of Alamar Blue was added to eachwell of the black 384-well plates. The plates were incubated for 2 hoursin the incubator (37° C.+5% CO₂) and at room temperature for one hour.Fluorescence of each plate was read on a plated reader for the AlamarBlue viability assay.

One set of white assay plates was warmed to room temperature (45minute).

The One Glo luciferase mixture was prepared. The media from white384-well assay plates was aspirated. Twenty-five μL of the One Gloluciferase mixture was added to each well of the assay plates. Theplates were incubated on the bench (room temperature) for 45 minutes,including 10 minutes of shaking at 700 rpm. The luminescence of eachplate was read on a plate reader.

1:100 DLR substrate and 1:100 LgBiT Protein dilution were added to thePromega Stop and Glo buffer and mixed just before addition to assayplates. Twenty-five microliters of Stop and Glo mixture was added toeach well. Assay plates incubated for at least 45 minutes, including 10minutes of shaking at 700 rpm. The luminescence of each plate was readon a plate reader.

Analysis of LRRK2 HiBit Screening assays. As mentioned above, plateswere run in triplicate and the assay repeated twice (total of 6replicate for exemplary compound). For each treatment, measurements weretaken for firefly luciferase for cell number, cell viability (AlamarBlue), and Nanoluc for the LRRK2-HiBit quantification.

The LRRK2 HiBit and alamar blue signal was normalized to % DMSO medianvalue for each plate. Curve fitting was performed on each compound forreplicates across three plates. The date for exemplary compounds ofTable 1 below is shown below in Table 2 in the columns labeled *WT DC50(nM) and **WT Dmax (%).

TABLE 1 Exemplary bifunctional compounds of the present disclosure. Ex.Structure Mass Name 1

949.48 N-(3-(2-((3S,5R)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3,5-dimethylpiperazin-1-yl)pyridin- 4-yl)-1H-indazol-5-yl)-4,5-dimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 2

963.50 N-(3-(2-((3S,5R)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3,5-dimethylpiperazin-1-yl)pyridin- 4-yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 3

911.43 N-(3-(2-((3R,5S)-4-(2-(2-(4-(2- (2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1- yl)ethoxy)ethyl)-3,5-dimethylpiperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5-dimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 4

955.46 N-(3-(2-((3R,5S)-4-(2-(2-(2-(4-(2- (2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1- yl)ethoxy)ethoxy)ethyl)-3,5-dimethylpiperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5-dimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 5

867.40 N-(3-(2-((3R,5S)-4-(2-(4-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1- yl)ethyl)-3,5-dimethylpiperazin-1-yl)pyridin-4-yl)-1H-indazol-5-yl)- 4,5-dimethyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 6

977.51 N-(3-(2-((3S,5R)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3,5-dimethylpiperazin-1-yl)pyridin- 4-yl)-1H-indazol-5-yl)-4,5,7,7-tetramethyl-4,7- dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 7

963.50 (S)-N-(3-(2-((3S,5R)-4-((1-((1-(2- (2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3,5-dimethylpiperazin-1-yl)pyridin- 4-yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 8

949.48 (R)-N-(3-(2-((S)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3-methylpiperazin-1-yl)pyridin-4-yl)- 1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 9

949.48 (S)-N-(3-(2-((S)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3-methylpiperazin-1-yl)pyridin-4-yl)- 1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 10

935.47 N-(3-(2-((S)-4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)-3-methylpiperazin-1-yl)pyridin-4-yl)- 1H-indazol-5-yl)-4,5-dimethyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 11

935.47 (7R)-N-(3-(2-(4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 12

935.47 (7S)-N-(3-(2-(4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 13

921.45 N-(3-(2-(4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)methyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5-dimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 14

949.48 (7R)-N-(3-(2-(4-(2-(1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)ethyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 15

867.40 (7R)-N-(3-(2-(4-(3-(4-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1- yl)propyl)piperazin-1-yl)pyridin-4-yl)-1H-indazol-5-yl)-4,5,7- trimethyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine-6-carboxamide 16

893.42 (7R)-N-(3-(2-(4-((1-(1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)azetidin-3-yl)methyl)piperazin-1-yl)pyridin-4-yl)-1H-indazol-5-yl)- 4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 17

907.44 (7R)-N-(3-(2-(4-((1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)azetidin-3-yl)methyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 18

921.45 (7R)-N-(3-(2-(4-((1-(1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)piperidin-4-yl)methyl)piperazin-1-yl)pyridin-4-yl)-1H-indazol-5-yl)- 4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 19

850.38 20

909.41 (7R)-N-[3-(2-{4-[2-(2-{6-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]-2,6-diazaspiro[3.3]heptan-2- yl}ethoxy)ethyl]piperazin-1-yl}pyridin-4-yl)-1H-indazol-5-yl]- 4,5,7-trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 21

950.48 (7R)-N-(3-(2-(4-(2-(4-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperazin-1-yl)ethyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-y1)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 22

985.46 (7R)-N-(3-(2-(4-(2-(1-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-4-yl)-2,2-difluoroethyl)piperazin-1- yl)pyridin-4-yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7- dihydrotetrazolo[1,5-a]pyrimidine- 6-carboxamide 23

949.48 (7R)-N-(3-(2-(4-(2-(4-((1-(2-(2,6- dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4- yl)methyl)piperidin-1-yl)ethyl)piperazin-1-yl)pyridin-4- yl)-1H-indazol-5-yl)-4,5,7-trimethyl-4,7-dihydrotetrazolo[1,5- a]pyrimidine-6-carboxamide 24

893.42 (7R)-N-[3-(2-{4-[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperidin-4-yl} methyl)azetidin-3- yl]piperazin-1-yl}pyridin-4-yl)-1H-indazol-5-yl]-4,5,7-trimethyl- 4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide 25

950.48 (7R)-N-{3-[2-(4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1- yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7- trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 26

850.38 (7R)-N-(3-{2-[6-({1-[2-(2,6- dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin- 4-yl}methyl)-2,6-diazaspiro[3.3]heptan-2-yl]pyridin- 4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 27

950.48 (7S)-N-{3-[2-(4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1- yl]ethyl}piperazin-1-yl)pyridin-4-yl]-1H-indazol-5-yl}-4,5,7- trimethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 28

964.49 (7S)-N-(3-{2-[(3S)-4-{2-[4-({1-[2- (2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5- yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}-3-methylpiperazin-1- yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide 29

949.48 N-{3-[2-(4-{[1-({1-[2-(2,6- dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin- 4-yl}methyl)piperidin-4-yl]methyl}piperazin-1-yl)pyridin-4- yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 30

978.51 N-(3-{2-[(3S)-4-{2-[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1-yl]ethyl}- 3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7- tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 31

964.49 (7S)-N-(3-{2-[(3S)-4-{2-[4-({4-[2- (2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5- yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}-3-methylpiperazin-1- yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7-trimethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a]pyrimidine-6-carboxamide 32

951.46 N-(3-{2-[(2S)-2-{[4-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperazin-1- yl]methyl}morpholin-4-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7- tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 33

964.49 N-{3-[2-(4-{2-[4-({4-[2-(2,6- dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperazin- 1-yl}methyl)piperidin-1-yl]ethyl}piperazin-1-yl)pyridin-4- yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 34

963.50 N-(3-{2-[(3S)-4-{[1-({1-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperidin-4-yl}methyl)piperidin-4- yl]methyl}-3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)- 4,5,7,7-tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide 35

964.49 N-{3-[2-(4-{2-[4-({1-[2-(2,6- dioxopiperidin-3-yl)-1,3-dioxo-2,3-dihydro-1H-isoindol-5-yl]piperidin- 4-yl}methyl)piperazin-1-yl]ethyl}piperazin-1-yl)pyridin-4- yl]-1H-indazol-5-yl}-4,5,7,7-tetramethyl-4H,7H- [1,2,3,4]tetrazolo[1,5-a ]pyrimidine- 6-carboxamide36

978.51 N-(3-{2-[(3S)-4-{2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo-2,3- dihydro-1H-isoindol-5-yl]piperazin-1-yl}methyl)piperidin-1-yl]ethyl}- 3-methylpiperazin-1-yl]pyridin-4-yl}-1H-indazol-5-yl)-4,5,7,7- tetramethyl-4H,7H-[1,2,3,4]tetrazolo[1,5-a]pyrimidine- 6-carboxamide

TABLE 2 Characterization of exemplary bifunctional compounds of thepresent disclosure. Mean *G2019S **G2019 *WT ** WT Observed DC50 S DmaxDC50 Dmax Ex. NMR Mass (nM) (%) (nM) (%)  1 ¹H NMR: (400 MHz, DMSO-d₆)δ: 13.42 (s, 1H), 11.08 (s, 1H), 10.07 (s, 1H), 8.67 950.60 A B B B (s,1H), 8.22 (d, J = 5.1 Hz, 1H), 7.67-7.51 (m, 3H), 7.28 (d, J = 13.8 Hz,2H), 7.23 (d, J = 8.8 Hz, 1H), 7.16 (d, J = 5.3 Hz, 1H), 5.30 (s, 2H),5.11-5.02 (m, 1H), 4.12 (d, J = 11.1 Hz, 2H), 4.03 (d, J = 12.9 Hz, 2H),3.44 (s, 3H), 3.00-2.80 (m, 5H), 2.72- 2.53 (m, 6H), 2.39-2.30 (m, 2H),2.27 (s, 3H), 2.16 (d, J = 5.8 Hz, 2H), 2.05-1.97 (m, 1H), 1.95-1.71 (m,7H), 1.37 (s, 1H), 1.12 (d, J = 5.8 Hz, 10H).  2 ¹H NMR: (400 MHz, DMSO)δ: 13.42 (s, 1H), 11.08 (s, 1H), 10.28 (s, 1H), 8.67 (s, 964.60 A B B B1H), 8.23 (d, J = 5.3 Hz, 1H), 8.15 (s, 1H), 7.67-7.58 (m, 2H),7.56-7.50 (m, 1H), 7.28 (d, J = 13.6 Hz, 2H), 7.23 (d, J = 8.8 Hz, 1H),7.16 (d, J = 5.4 Hz, 1H), 5.83- 5.71 (m, 1H), 5.06 (dd, J = 5.4, 12.9Hz, 1H), 4.12 (d, J = 12.0 Hz, 2H), 4.03 (d, J = 12.9 Hz, 2H), 3.44 (s,3H), 3.01-2.82 (m, 5H), 2.72-2.55 (m, 5H), 2.40-2.33 (m, 2H), 2.20 (s,3H), 2.17 (s, 2H), 2.06-1.97 (m, 1H), 1.94-1.71 (m, 7H), 1.56 (d, J =6.4 Hz, 3H), 1.38 (s, 1H), 1.11 (d, J = 5.8 Hz, 10H).  3 ¹H NMR: (400MHz, CD₃OD) δ: 8.65 (s, 1H), 8.40 (s, 1H), 8.17 (d, J = 5.3 Hz, 1H),912.57 D — D — 7.52 (d, J = 8.9 Hz, 1H), 7.38-7.32 (m, 2H), 7.24 (d, J =5.3 Hz, 1H), 7.19 (d, J = 8.5 Hz, 1H), 6.93 (d, J = 2.1 Hz, 1H), 6.75(dd, J = 2.1, 8.5 Hz, 1H), 5.30 (s, 2H), 5.02 (br dd, J = 5.5, 12.8 Hz,1H), 4.53 (br t, J = 11.4 Hz, 2H), 3.92-3.81 (m, 4H), 3.67 (br t, J =4.8 Hz, 2H), 3.55 (br s, 2H), 3.51 (s, 3H), 3.26-3.20 (m, 4H), 3.04-2.94(m, 2H), 2.92-2.81 (m, 1H), 2.76 (br d, J = 2.8 Hz, 1H), 2.68-2.56 (m,7H), 2.36 (s, 3H), 2.14-2.05 (m, 1H), 1.48 (br d, J = 5.4 Hz, 6H)  4 ¹HNMR: (400 MHz, MeOD) δ: 8.71 (s, 1H), 8.27 (d, J = 5.4 Hz, 1H),7.55-7.48 956.59 D — D — (m, 2H), 7.41 (d, J = 8.5 Hz, 1H), 7.38-7.31(m, 2H), 6.99 (d, J = 1.8 Hz, 1H), 6.87 (d, J = 8.5 Hz, 1H), 5.29 (s,2H), 5.05 (dd, J = 5.4, 12.7 Hz, 1H), 4.55 (d, J = 13.9 Hz, 2H), 3.85(d, J = 4.5 Hz, 2H), 3.79-3.53 (m, 10H), 3.51 (s, 3H), 3.22 (s, 4H),3.05- 2.93 (m, 2H), 2.91-2.51 (m, 9H), 2.34 (s, 3H), 2.16-2.07 (m, 1H),1.49 (d, J = 6.3 Hz, 6H).  5 ¹H NMR: (400 MHz, METHANOL-d₄) δ: 8.69 (s,1H), 8.41 (br s, 1H), 8.28 (d, 868.54 D C D — J = 5.3 Hz, 1H), 7.54-7.47(m, 2H), 7.45 (s, 1H), 7.36 (d, J = 6.6 Hz, 2H), 7.15 (s, 1H), 7.01 (brd, J = 6.9 Hz, 1H), 5.29 (s, 2H), 5.08 (dd, J = 5.5, 12.4 Hz, 1H), 4.59(s, 3H), 4.43 (br s, 2H), 3.51 (s, 4H), 3.27 (br s, 4H), 3.02 (br t, J =12.6 Hz, 2H), 2.93- 2.82 (m, 1H), 2.80-2.75 (m, 1H), 2.75-2.69 (m, 3H),2.66 (br t, J = 4.7 Hz, 4H), 2.35 (s, 3H), 2.19-2.05 (m, 1H), 1.44 (brd, J = 4.9 Hz, 6H)  6 1H NMR (400 MHz, MeOD-d4) δ: 8.74 (s, 1H), 8.46(s, 1H), 8.15 (d, J = 5.4 Hz, 978.8 B C D C 1H), 7.68 (d, J = 8.5 Hz,1H), 7.58 (d, J = 8.9 Hz, 1H), 7.36 (d, J = 2.1 Hz, 1H), 7.31- 7.20 (m,4H), 5.07 (dd, J = 5.4, 12.3 Hz, 1H), 4.75 (br s, 1H), 4.71 (br s, 1H),4.14- 4.00 (m, 5H), 3.73 (s, 1H), 3.59 (br d, J = 11.0 Hz, 1H), 3.46 (s,3H), 3.03 (br d, J = 14.3 Hz, 5H), 2.89-2.81 (m, 4H), 2.77 (br s, 2H),2.74-2.67 (m, 2H), 2.63 (br d, J = 5.8 Hz, 2H), 2.20 (br d, J = 17.6 Hz,2H), 2.14 (br s, 2H), 1.97-1.91 (m, 2H), 1.88 (s, 3H), 1.57 (br d, J =14.9 Hz, 2H), 1.50 (s, 3H), 1.45-1.37 (m, 2H), 1.33- 1.25 (m, 2H),1.19-1.14 (m, 5H)  8 1H NMR (400 MHz, MeOD-d4)δ: 8.68 (s, 1H), 8.47 (brs, 1H), 8.22 (d, J = 5.6 Hz, 950.4 A B B B 1H), 7.68 (d, J = 8.5 Hz,1H), 7.61 (d, J = 9.0 Hz, 1H), 7.50 (d, J = 8.6 Hz, 1H), 7.38 (br d, J =5.9 Hz, 2H), 7.31-7.21 (m, 2H), 5.74 (br d, J = 5.8 Hz, 1H), 5.07 (dd, J= 5.4, 12.1 Hz, 1H), 4.12-4.07 (m, 2H), 4.01 (br d, J = 9.5 Hz, 2H),3.50 (s, 5H), 3.11-2.99 (m, 5H), 2.95 (br s, 2H), 2.89-2.80 (m, 2H),2.79-2.69 (m, 3H), 2.62 (br s, 1H), 2.43 (br s, 1H), 2.28 (s, 3H),2.24-2.04 (m, 5H), 1.99-1.84 (m, 4H), 1.69 (d, J = 6.4 Hz, 3H),1.51-1.37 (m, 4H), 1.22-1.17 (m, 3H)  9 1H NMR (400 MHz, MeOD-d4) δ:8.68 (s, 1H), 8.42 (br s, 1H), 8.22 (d, J = 5.3 Hz, 950.4 C B D C 1H),7.68 (d, J = 8.5 Hz, 1H), 7.61 (d, J = 9.0 Hz, 1H), 7.49 (dd, J = 1.6,8.9 Hz, 1H), 7.41-7.34 (m, 2H), 7.30-7.21 (m, 2H), 5.74 (br d, J = 6.1Hz, 1H), 5.07 (dd, J = 5.4, 12.4 Hz, 1H), 4.09 (br d, J = 12.9 Hz, 2H),4.01 (br d, J = 12.3 Hz, 2H), 3.56 (br d, J = 9.0 Hz, 2H), 3.50 (s, 3H),3.10-2.97 (m, 7H), 2.96-2.81 (m, 3H), 2.78- 2.69 (m, 3H), 2.63 (br s,1H), 2.44 (br t, J = 9.7 Hz, 1H), 2.28 (s, 3H), 2.25-2.15 (m, 3H),2.13-2.06 (m, 1H), 1.99 (br d, J = 14.4 Hz, 1H), 1.92 (br d, J = 12.9Hz, 3H), 1.68 (d, J = 6.4 Hz, 3H), 1.56-1.36 (m, 4H), 1.20 (d, J = 6.1Hz, 3H) 10 1H NMR (400 MHz, MeOD-d4) δ: 8.64 (s, 1H), 8.44 (s, 2H), 8.22(d, J = 5.5 Hz, 936.5 A B B B 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.63-7.58(m, 1H), 7.50 (d, J = 9.1 Hz, 1H), 7.40- 7.34 (m, 2H), 7.31-7.22 (m,2H), 5.29 (s, 2H), 5.07 (dd, J = 5.4, 12.6 Hz, 1H), 4.60 (br s, 2H),4.09 (br d, J = 11.6 Hz, 2H), 4.01 (br d, J = 11.9 Hz, 2H), 3.55 (br s,2H), 3.51 (s, 3H), 3.12-2.96 (m, 5H), 2.96-2.81 (m, 3H), 2.78-2.68 (m,3H), 2.62 (br s, 1H), 2.43 (br t, J = 9.4 Hz, 1H), 2.34 (s, 3H), 2.20(br s, 3H), 2.12-2.07 (m, 1H), 2.02-1.89 (m, 4H), 1.57-1.35 (m, 4H),1.20 (d, J = 6.1 Hz, 3H) 11 1H NMR (400 MHz, MeOD-d4) δ: 8.64 (s, 1H),8.23 (d, J = 5.1 Hz, 1H), 7.63 (dd, 936.3 B C C C J = 8.6, 15.1 Hz, 2H),7.55-7.49 (m, 1H), 7.38 (s, 1H), 7.30 (d, J = 5.1 Hz, 1H), 6.99 (s, 1H),6.83 (d, J = 7.4 Hz, 1H), 5.74 (d, J = 6.0 Hz, 1H), 5.06 (dd, J = 5.5,12.8 Hz, 1H), 3.72-3.56 (m, 9H), 3.49 (s, 3H), 3.45 (s, 1H), 3.22-3.11(m, 4H), 2.97 (s, 3H), 2.89-2.64 (m, 7H), 2.45-2.33 (m, 2H), 2.28 (s,3H), 2.11 (d, J = 13.3 Hz, 3H), 1.94 (d, J = 7.4 Hz, 3H), 1.87-1.79 (m,1H), 1.68 (d, J = 6.3 Hz, 3H), 1.52 (d, J = 6.9 Hz, 1H) 12 1H NMR (400MHz, MeOD-d4) δ: 8.64 (s, 1H), 8.23 (d, J = 5.1 Hz, 1H), 7.63 (dd, 936.4D C D J = 8.6, 15.1 Hz, 2H), 7.55-7.49 (m, 1H), 7.38 (s, 1H), 7.30 (d, J= 5.1 Hz, 1H), 6.99 (s, 1H), 6.83 (d, J = 7.4 Hz, 1H), 5.74 (d, J = 6.0Hz, 1H), 5.06 (dd, J = 5.5, 12.8 Hz, 1H), 3.72-3.56 (m, 9H), 3.49 (s,3H), 3.45 (s, 1H), 3.22-3.11 (m, 4H), 2.97 (s, 3H), 2.89-2.64 (m, 7H),2.45-2.33 (m, 2H), 2.28 (s, 3H), 2.11 (d, J = 13.3 Hz, 3H), 1.94 (d, J =7.4 Hz, 3H), 1.87-1.79 (m, 1H), 1.68 (d, J = 6.3 Hz, 3H), 1.52 (d, J =6.9 Hz, 1H) 13 1H NMR (400 MHz, MeOD-d4) δ: 8.64 (s, 1H), 8.23 (d, J =5.1 Hz, 1H), 7.63 (dd, 922.4 B B D C J = 8.6, 15.1 Hz, 2H), 7.55-7.49(m, 1H), 7.38 (s, 1H), 7.30 (d, J = 5.1 Hz, 1H), 6.99 (s, 1H), 6.83 (d,J = 7.4 Hz, 1H), 5.74 (d, J = 6.0 Hz, 1H), 5.06 (dd, J = 5.5, 12.8 Hz,1H), 3.72-3.56 (m, 9H), 3.49 (s, 3H), 3.45 (s, 1H), 3.22-3.11 (m, 4H),2.97 (s, 3H), 2.89-2.64 (m, 7H), 2.45-2.33 (m, 2H), 2.28 (s, 3H), 2.11(d, J = 13.3 Hz, 3H), 1.94 (d, J = 7.4 Hz, 3H), 1.87-1.79 (m, 1H), 1.68(d, J = 6.3 Hz, 3H), 1.52 (d, J = 6.9 Hz, 1H) 14 1H NMR (400 MHz,MeOD-d4) δ: 8.75-8.61 (m, 1H), 8.38 (s, 2H), 8.28-8.18 950.4 C C D B (m,1H), 7.63 (dd, J = 8.7, 13.6 Hz, 2H), 7.57-7.49 (m, 1H), 7.41 (s, 1H),7.35- 7.28 (m, 1H), 6.98 (s, 1H), 6.87-6.80 (m, 1H), 5.74 (br d, J = 6.0Hz, 1H), 5.06 (dd, J = 5.6, 12.5 Hz, 1H), 4.62 (br s, 1H), 3.80-3.65 (m,5H), 3.57 (br d, J = 9.0 Hz, 3H), 3.51-3.42 (m, 4H), 3.23-3.10 (m, 3H),3.06-2.92 (m, 2H), 2.88-2.79 (m, 5H), 2.74 (br d, J = 15.9 Hz, 2H),2.70-2.62 (m, 2H), 2.41 (br s, 1H), 2.28 (s, 3H), 2.15-2.01 (m, 3H),1.93 (br d, J = 8.4 Hz, 2H), 1.87-1.73 (m, 2H), 1.70-1.61 (m, 5H), 1.52(br d, J = 6.5 Hz, 2H) 15 1H NMR (400 MHz, MeOD-d4) δ: 8.67 (s, 1H),8.28 (d, J = 5.5 Hz, 1H), 7.69 (d, 868.3 B B D B J = 8.5 Hz, 1H),7.64-7.60 (m, 1H), 7.50 (dd, J = 1.8, 8.9 Hz, 1H), 7.45 (s, 1H), 7.39-7.33 (m, 2H), 7.26 (dd, J = 2.3, 8.4 Hz, 1H), 5.74 (d, J = 5.8 Hz, 1H),5.07 (dd, J = 5.5, 12.4 Hz, 1H), 3.85 (s, 4H), 3.57-3.52 (m, 4H),3.51-3.47 (m, 3H), 3.21- 3.13 (m, 4H), 3.02 (t, J = 7.2 Hz, 2H),2.88-2.80 (m, 5H), 2.78-2.69 (m, 4H), 2.28 (d, J = 0.9 Hz, 3H),2.14-2.07 (m, 1H), 2.04-1.97 (m, 2H), 1.68 (d, J = 6.4 Hz, 3H) 17 1H NMR(400 MHz, MeOD-d4) δ: 8.56 (d, J = 8.9 Hz, 1H), 8.23 (d, J = 5.3 Hz,1H), 894.6 C B D B 7.66 (d, J = 8.5 Hz, 1H), 7.63-7.54 (m, 2H), 7.37 (s,1H), 7.35 (d, J = 2.1 Hz, 1H), 7.30 (d, J = 5.4 Hz, 1H), 7.22 (dd, J =2.3, 8.6 Hz, 1H), 5.74 (q, J = 6.0 Hz, 1H), 5.06 (dd, J = 5.4, 12.6 Hz,1H), 4.02 (br d, J = 13.3 Hz, 2H), 3.68-3.62 (m, 4H), 3.59 (br t, J =7.6 Hz, 2H), 3.50 (s, 3H), 3.02-2.97 (m, 3H), 2.91-2.79 (m, 2H),2.78-2.69 (m, 2H), 2.68-2.59 (m, 6H), 2.42 (br t, J = 10.4 Hz, 1H), 2.28(d, J = 1.0 Hz, 3H), 2.15-2.07 (m, 1H), 1.88 (br d, J = 12.6 Hz, 2H),1.69 (d, J = 6.4 Hz, 3H), 1.29 (br s, 3H) 18 1H NMR (400 MHz, MeOD-d4)δ: 8.65 (s, 1H), 8.29 (br s, 2H), 8.23 (d, J = 5.4 Hz, 922.6 C B D C1H), 7.72 (d, J = 8.5 Hz, 1H), 7.64-7.59 (m, 1H), 7.51 (dd, J = 1.8, 9.0Hz, 1H), 7.42 (d, J = 2.1 Hz, 1H), 7.39 (s, 1H), 7.33-7.27 (m, 2H),5.82-5.69 (m, 1H), 5.15- 5.02 (m, 1H), 4.24 (br d, J = 13.5 Hz, 2H),3.66 (br s, 4H), 3.58 (br d, J = 11.6 Hz, 2H), 3.50 (s, 3H), 3.13-2.99(m, 4H), 2.94-2.81 (m, 1H), 2.79-2.58 (m, 6H), 2.56 (s, 1H), 2.38 (br d,J = 7.1 Hz, 1H), 2.28 (s, 3H), 2.26-2.07 (m, 5H), 1.99 (br s, 1H),1.90-1.76 (m, 2H), 1.68 (d, J = 6.4 Hz, 3H), 1.57-1.42 (m, 3H) 20 1H NMR(400 MHz, MeOD-d4) δ: 8.67 (s, 1H), 8.34 (br s, 2H), 8.27 (d, J = 5.3Hz, 910.5 D D 1H), 7.57 (dd, J = 8.6, 18.2 Hz, 2H), 7.46 (dd, J = 1.6,9.0 Hz, 1H), 7.42 (s, 1H), 7.34 (d, J = 5.3 Hz, 1H), 6.76 (d, J = 1.9Hz, 1H), 6.60 (br d, J = 8.4 Hz, 1H), 5.74 (br d, J = 5.8 Hz, 1H), 5.04(dd, J = 5.4, 12.7 Hz, 1H), 4.22 (s, 8H), 3.82-3.71 (m, 6H), 3.67 (br t,J = 4.9 Hz, 2H), 3.48 (s, 3H), 3.28 (br s, 1H), 2.94-2.80 (m, 7H), 2.79-2.61 (m, 3H), 2.26 (s, 3H), 2.13-2.03 (m, 1H), 1.67 (d, J = 6.3 Hz, 3H)21 1H NMR (400 MHz, MeOD-d4) δ: 8.65 (s, 1H), 8.35 (s, 2H), 8.25 (d, J =5.4 Hz, 951.6 D B D 1H), 7.64 (dd, J = 8.9, 18.9 Hz, 2H), 7.53-7.47 (m,1H), 7.40 (s, 1H), 7.36-7.30 (m, 2H), 7.25-7.19 (m, 1H), 5.78-5.71 (m,1H), 5.06 (dd, J = 5.7, 12.7 Hz, 1H), 4.05 (br d, J = 13.5 Hz, 2H), 3.72(br s, 4H), 3.52-3.48 (m, 4H), 3.11-2.96 (m, 7H), 2.90-2.70 (m, 12H),2.43 (br d, J = 4.4 Hz, 2H), 2.28 (d, J = 1.0 Hz, 3H), 2.11 (br d, J =5.3 Hz, 1H), 1.91 (br d, J = 9.8 Hz, 3H), 1.69 (d, J = 6.4 Hz, 3H), 1.31(br d, J = 14.6 Hz, 3H) 22 1H NMR (400 MHz, MeOD-d4) δ: 8.61 (s, 1H),8.48 (br s, 1H), 8.23 (d, J = 5.3 Hz, 986.4 A B A B 1H), 7.67 (d, J =8.5 Hz, 1H), 7.63-7.59 (m, 1H), 7.55-7.50 (m, 1H), 7.39-7.34 (m, 2H),7.30 (d, J = 5.1 Hz, 1H), 7.22 (dd, J = 2.3, 8.6 Hz, 1H), 5.74 (br d, J= 6.1 Hz, 1H), 5.19 (s, 1H), 4.11-4.03 (m, 2H), 3.65 (br s, 4H), 3.50(s, 3H), 3.42-3.37 (m, 1H), 3.02 (br t, J = 11.8 Hz, 2H), 2.92-2.80 (m,4H), 2.79-2.76 (m, 4H), 2.74- 2.64 (m, 4H), 2.55 (br s, 1H), 2.33 (br d,J = 7.6 Hz, 1H), 2.28 (d, J = 1.0 Hz, 3H), 2.15-2.05 (m, 2H), 2.03-1.96(m, 2H), 1.91 (br d, J = 11.5 Hz, 2H), 1.83-1.74 (m, 2H), 1.68 (d, J =6.3 Hz, 3H), 1.44-1.27 (m, 3H) 23 1H NMR (400 MHz, MeOD-d4) δ: 8.69 (s,1H), 8.44 (s, 1H), 8.27 (d, J = 5.3 Hz, 950.6 B B C B 1H), 7.72-7.58 (m,2H), 7.52 (dd, J = 1.7, 8.9 Hz, 1H), 7.42 (s, 1H), 7.37-7.31 (m, 2H),7.22 (dd, J = 2.3, 8.6 Hz, 1H), 5.76 (br d, J = 6.6 Hz, 1H), 5.08 (dd, J= 5.5, 12.5 Hz, 1H), 4.06 (br d, J = 13.1 Hz, 2H), 3.71 (br s, 4H), 3.64(br d, J = 15.3 Hz, 2H), 3.52 (s, 3H), 3.11-2.95 (m, 5H), 2.87-2.69 (m,11H), 2.30 (s, 3H), 2.17-2.09 (m, 1H), 2.01 (br d, J = 12.9 Hz, 2H),1.86 (br d, J = 11.9 Hz, 2H), 1.70 (d, J = 6.4 Hz, 3H), 1.56-1.45 (m,2H), 1.38-1.23 (m, 5H) 24 1H NMR (400 MHz, MeOD-d4) δ: 8.65 (s, 1H),8.47 (br s, 1H), 8.24 (d, J = 5.3 Hz, 894.5 B B B C 1H), 7.67 (d, J =8.5 Hz, 1H), 7.64-7.59 (m, 1H), 7.52 (d, J = 9.0 Hz, 1H), 7.41- 7.34 (m,2H), 7.32 (d, J = 5.3 Hz, 1H), 7.23 (dd, J = 2.1, 8.6 Hz, 1H), 5.78-5.70(m, 1H), 5.07 (dd, J = 5.4, 12.6 Hz, 1H), 4.08 (br d, J = 12.9 Hz, 4H),3.82 (br s, 2H), 3.69 (br s, 4H), 3.50 (s, 3H), 3.27 (br s, 1H),3.07-2.96 (m, 4H), 2.92-2.81 (m, 1H), 2.78-2.63 (m, 2H), 2.57 (br s,4H), 2.28 (s, 3H), 2.15-2.06 (m, 1H), 1.95- 1.81 (m, 3H), 1.68 (d, J =6.3 Hz, 3H), 1.44-1.32 (m, 2H) 25 1H NMR (400 MHz, MeOD-d4) δ: 8.64 (s,1H), 8.45 (s, 1H), 8.25 (d, J = 5.3 Hz, 951.6 B A C A 1H), 7.66 (d, J =8.6 Hz, 1H), 7.63-7.60 (m, 1H), 7.52 (dd, J = 1.7, 8.9 Hz, 1H), 7.40 (s,1H), 7.35-7.30 (m, 2H), 7.21 (dd, J = 2.2, 8.6 Hz, 1H), 5.74 (br d, J =5.9 Hz, 1H), 5.06 (br dd, J = 5.4, 12.5 Hz, 1H), 4.05 (br d, J = 13.5Hz, 2H), 3.71 (br s, 4H), 3.50 (s, 3H), 3.00 (br t, J = 11.4 Hz, 7H),2.88-2.67 (m, 13H), 2.40 (br d, J = 6.3 Hz, 2H), 2.28 (s, 3H), 2.15-2.06(m, 1H), 1.90 (br d, J = 10.9 Hz, 3H), 1.68 (d, J = 6.4 Hz, 3H),1.37-1.25 (m, 3H) 26 1H NMR: (400 MHz, DMSO) δ: 13.45 (s, 1H), 11.07 (s,1H), 10.29 (s, 1H), 8.53 851.3 D A D (s, 1H), 8.21-8.18 (m, 1H),7.66-7.58 (m, 3H), 7.29 (s, 1H), 7.24-7.17 (m, 2H), 6.86-6.79 (m, 1H),5.80-5.68 (m, 1H), 5.06 (dd, J = 5.4, 12.9 Hz, 1H), 4.08 (s, 3H), 4.02(d, J = 13.3 Hz, 2H), 3.35-3.27 (m, 8H), 2.99-2.79 (m, 4H), 2.69-2.53(m, 3H), 2.33 (s, 1H), 2.20 (s, 3H), 2.07-1.97 (m, 1H), 1.75 (d, J =11.5 Hz, 2H), 1.57 (d, J = 6.4 Hz, 3H), 1.23-1.11 (m, 2H) 27 1H NMR (400MHz, MeOD-d4) δ: 8.67 (s, 1H), 8.26 (d, J = 5.3 Hz, 1H), 7.70 (d, 951.3D B D J = 8.5 Hz, 1H), 7.62 (d, J = 8.9 Hz, 1H), 7.50 (d, J = 1.8 Hz,1H), 7.40 (s, 1H), 7.37 (d, J = 2.1 Hz, 1H), 7.32 (d, J = 5.5 Hz, 1H),7.24 (dd, J = 2.3, 8.6 Hz, 1H), 5.75 (d, J = 5.8 Hz, 1H), 5.08 (dd, J =5.5, 12.5 Hz, 1H), 3.70 (d, J = 4.6 Hz, 4H), 3.67-3.59 (m, 3H), 3.51 (s,3H), 3.48 (d, J = 3.8 Hz, 4H), 3.10-3.02 (m, 2H), 2.90-2.81 (m, 3H),2.80-2.72 (m, 6H), 2.68-2.59 (m, 5H), 2.35 (d, J = 7.3 Hz, 2H), 2.29 (d,J = 0.9 Hz, 3H), 2.16-2.06 (m, 3H), 1.95 (s, 1H), 1.69(d, J = 6.3 Hz,3H), 1.59- 1.48 (m, 2H) 28 1H NMR (400 MHz, MeOD-d4) δ: 8.72 (s, 1H),8.31 (s, 2H), 8.25 (d, J = 5.4 Hz, 965.7 D B D 1H), 7.69-7.54 (m, 2H),7.48 (dd, J = 1.8, 8.9 Hz, 1H), 7.43 (s, 1H), 7.37-7.29 (m, 2H), 7.21(dd, J = 2.3, 8.7 Hz, 1H), 5.74 (br d, J = 5.9 Hz, 1H), 5.06 (dd, J =5.4, 12.4 Hz, 1H), 4.06 (br t, J = 13.4 Hz, 4H), 3.52-3.49 (m, 3H),3.27-3.15 (m, 4H), 3.11- 2.90 (m, 9H), 2.87-2.66 (m, 9H), 2.42 (br d, J= 6.3 Hz, 2H), 2.28 (d, J = 0.9 Hz, 3H), 2.16-2.05 (m, 1H), 1.95-1.83(m, 3H), 1.68 (d, J = 6.4 Hz, 3H), 1.34-1.24 (m, 5H) 29 1H NMR (400 MHz,MeOD-d4) δ: 8.78 (s, 1H), 8.47 (s, 2H), 8.17 (d, J = 5.3 Hz, 950.6 B C DC 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 9.0 Hz, 1H), 7.37-7.32 (m,2H), 7.29- 7.22 (m, 3H), 5.07 (dd, J = 5.4, 12.4 Hz, 1H), 4.75 (br s,1H), 4.71 (s, 1H), 4.09 (br d, J = 13.3 Hz, 2H), 3.70 (s, 1H), 3.63 (brd, J = 11.6 Hz, 2H), 3.58 (br s, 4H), 3.48 (s, 3H), 3.16 (br s, 1H),3.08-3.01 (m, 4H), 2.88-2.82 (m, 1H), 2.79-2.72 (m, 2H), 2.68 (br d, J =13.1 Hz, 1H), 2.60 (br s, 3H), 2.43 (br d, J = 7.0 Hz, 2H), 2.23- 2.17(m, 3H), 2.13-2.07 (m, 1H), 1.98-1.86 (m, 6H), 1.61 (br d, J = 11.4 Hz,2H), 1.51 (s, 3H), 1.46-1.39 (m, 2H), 1.33-1.28 (m, 1H) 30 1H NMR (400MHz, MeOD-d4) δ: 8.85 (s, 1H), 8.39 (s, 2H), 8.18 (d, J = 5.4 Hz, 979.7B C D C 1H), 7.65 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 8.9 Hz, 1H),7.37-7.30 (m, 2H), 7.29- 7.22 (m, 2H), 7.19 (dd, J = 2.0, 8.6 Hz, 1H),5.06 (dd, J = 5.4, 12.4 Hz, 1H), 4.75 (br s, 1H), 4.71 (s, 1H),4.06-3.96 (m, 3H), 3.88-3.81 (m, 1H), 3.71 (s, 1H), 3.47 (d, J = 2.0 Hz,4H), 3.25 (br d, J = 10.8 Hz, 1H), 3.22-3.14 (m, 5H), 3.12 (br d, J =12.9 Hz, 1H), 3.01-2.93 (m, 3H), 2.90 (br d, J = 4.8 Hz, 1H), 2.88-2.85(m, 1H), 2.84- 2.74 (m, 6H), 2.74-2.64 (m, 2H), 2.39 (br d, J = 4.3 Hz,2H), 2.28-2.04 (m, 2H), 1.91-1.83 (m, 6H), 1.51 (s, 2H), 1.33-1.19 (m,6H) 31 1H NMR (400 MHz, MeOD-d4) δ: 8.73-8.64 (m, 1H), 8.49 (br s, 1H),8.23 (d, 965.8 D C D J = 5.5 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H),7.64-7.56 (m, 1H), 7.49 (br d, J = 9.0 Hz, 1H), 7.41-7.31 (m, 2H), 7.30(br d, J = 4.8 Hz, 1H), 7.26-7.19 (m, 1H), 5.73 (br d, J = 6.5 Hz, 1H),5.07 (br dd, J = 5.5, 12.3 Hz, 1H), 3.98 (br d, J = 12.6 Hz, 2H), 3.57-3.37 (m, 9H), 3.23-3.04 (m, 2H), 2.96-2.68 (m, 12H), 2.61 (br s, 4H),2.33 (br d, J = 7.4 Hz, 2H), 2.28 (s, 2H), 2.17-1.98 (m, 4H), 1.88 (brs, 1H), 1.68 (d, J = 6.4 Hz, 2H), 1.61-1.42 (m, 2H), 1.31-1.14 (m, 4H)32 1H NMR (400 MHz, MeOD-d4) δ: 8.54 (dd, J = 1.3, 8.9 Hz, 1H), 8.51 (s,1H), 8.22 952.6 B C D (d, J = 5.4 Hz, 1H), 7.65 (d, J = 8.5 Hz, 1H),7.57 (d, J = 8.9 Hz, 1H), 7.37-7.32 (m, 2H), 7.30 (s, 1H), 7.27 (d, J =5.3 Hz, 1H), 7.20 (dd, J = 2.3, 8.6 Hz, 1H), 5.06 (dd, J = 5.4, 12.4 Hz,1H), 4.75 (s, 1H), 4.70 (d, J = 1.5 Hz, 1H), 4.19 (br d, J = 12.5 Hz,1H), 4.08-4.00 (m, 4H), 3.88 (br s, 1H), 3.77-3.71 (m, 1H), 3.70 (s,1H), 3.47 (d, J = 0.8 Hz, 3H), 3.05-2.93 (m, 6H), 2.89-2.81 (m, 3H),2.81-2.75 (m, 4H), 2.74- 2.63 (m, 4H), 2.44 (br d, J = 6.0 Hz, 2H),2.14-2.07 (m, 1H), 1.92-1.87 (m, 6H), 1.51 (s, 3H), 1.32 (br s, 2H) 331H NMR (400 MHz, MeOD-d4) δ: 8.88 (s, 1H), 8.39 (s, 1H), 8.16 (d, J =5.4 Hz, 965.7 D C D C 1H), 7.68 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 8.9Hz, 1H), 7.38-7.33 (m, 2H), 7.30- 7.20 (m, 3H), 5.07 (br dd, J = 5.4,12.4 Hz, 1H), 4.76 (br s, 1H), 4.72 (br s, 1H), 3.82 (br d, J = 9.6 Hz,2H), 3.71 (s, 1H), 3.58 (br s, 3H), 3.54-3.51 (m, 1H), 3.50- 3.46 (m,6H), 3.21 (br t, J = 11.7 Hz, 2H), 2.88 (br t, J = 6.3 Hz, 2H), 2.83 (brd, J = 5.6 Hz, 1H), 2.78-2.75 (m, 1H), 2.73 (br s, 1H), 2.71-2.60 (m,9H), 2.41-2.33 (m, 2H), 2.22-2.06 (m, 4H), 2.03 (br d, J = 7.1 Hz, 1H),1.90-1.86 (m, 3H), 1.68- 1.57 (m, 2H), 1.52 (s, 3H), 1.34-1.27 (m, 1H)34 1H NMR (400 MHz, MeOD-d4) δ: 8.76 (br s, 1H), 8.53 (s, 1H), 8.15 (d,J = 5.3 Hz, 964.7 B C D 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 8.9Hz, 1H), 7.36 (d, J = 2.1 Hz, 1H), 7.31 (s, 1H), 7.29-7.19 (m, 3H), 5.07(dd, J = 5.5, 12.4 Hz, 1H), 4.75 (br s, 1H), 4.71 (br s, 1H), 4.09 (brd, J = 12.8 Hz, 2H), 3.92 (br d, J = 11.5 Hz, 1H), 3.80 (br d, J = 12.0Hz, 1H), 3.71 (s, 1H), 3.54 (br s, 2H), 3.47 (s, 3H), 3.07-2.97 (m, 6H),2.91-2.81 (m, 2H), 2.75-2.65 (m, 3H), 2.57 (br s, 1H), 2.28-2.19 (m,3H), 2.15-2.09 (m, 2H), 2.04 (br s, 1H), 1.93 (br d, J = 11.9 Hz, 2H),1.88 (s, 3H), 1.58 (br d, J = 8.6 Hz, 2H), 1.50 (s, 3H), 1.42 (br d, J =10.9 Hz, 2H), 1.35-1.27 (m, 4H), 1.11 (br t, J = 6.4 Hz, 3H) 35 1H NMR(400 MHz, MeOD-d4) δ: 8.83 (s, 1H), 8.45 (s, 1H), 8.18 (d, J = 5.3 Hz,965.6 B C D 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 8.9 Hz, 1H),7.35-7.31 (m, 2H), 7.29- 7.19 (m, 3H), 5.06 (br dd, J = 5.4, 12.3 Hz,1H), 4.75-4.75 (m, 1H), 4.71 (br s, 1H), 4.06 (br s, 1H), 4.03 (br s,1H), 3.70 (s, 1H), 3.62 (br s, 4H), 3.48 (s, 3H), 2.99 (br t, J = 12.0Hz, 4H), 2.90 (br s, 2H), 2.86 (br d, J = 4.1 Hz, 1H), 2.87-2.85 (m,1H), 2.82 (br s, 4H), 2.76 (br s, 2H), 2.73 (br s, 2H), 2.72-2.65 (m,1H), 2.70-2.65 (m, 1H), 2.41 (br d, J = 5.5 Hz, 2H), 2.12-2.06 (m, 1H),1.95-1.84 (m, 8H), 1.51 (s, 3H), 1.30 (br d, J = 11.4 Hz, 4H) 36 1H NMR(400 MHz, MeOD-d4) δ: 8.79 (br s, 1H), 8.55 (s, 1H), 8.16 (d, J = 5.4979.7 D C D Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 9.0 Hz, 1H),7.34 (br d, J = 12.4 Hz, 2H), 7.28-7.18 (m, 3H), 5.07 (br dd, J = 5.3,12.3 Hz, 1H), 4.75 (br s, 1H), 4.70 (br s, 1H), 3.90 (br d, J = 11.4 Hz,1H), 3.77 (br d, J = 11.6 Hz, 1H), 3.70 (s, 1H), 3.50-3.44 (m, 8H), 3.20(br d, J = 12.9 Hz, 2H), 2.99 (br d, J = 3.1 Hz, 1H), 2.91- 2.81 (m,3H), 2.71 (br d, J = 10.9 Hz, 4H), 2.60 (br s, 4H), 2.49 (br t, J = 9.1Hz, 1H), 2.33 (br d, J = 6.5 Hz, 2H), 2.24-2.13 (m, 1H), 2.13-2.08 (m,1H), 2.03 (br d, J = 12.9 Hz, 2H), 1.90-1.86 (m, 4H), 1.51 (s, 4H),1.36-1.23 (m, 4H), 1.16 (br t, J = 6.6 Hz, 3H) *DC₅₀ Ranges: A < 10; 10≤ B < 50; 50 ≤ C < 100; D ≥ 100. **D_(Max) Ranges: A ≥ 70; 50 ≤ B < 70;C < 50

A novel bifunctional molecule, which contains a recruiting moiety thatselectively or preferentially binds to a LRRK2 protein having at leastone mutation that is G2019S and an E3 ubiquitin ligase recruiting moietyis described. The bifunctional molecules of the present disclosureactively ubiquitinate the mutated LRRK2, resulting in proteasomaldegradation, leading to suppression of cellular proliferation andinduction of apoptosis.

The contents of all references, patents, pending patent applications andpublished patents, cited throughout this application are herebyexpressly incorporated by reference.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the disclosure described herein. Such equivalents areintended to be encompassed by the following claims. It is understoodthat the detailed examples and embodiments described herein are given byway of example for illustrative purposes only, and are in no wayconsidered to be limiting to the disclosure. Various modifications orchanges in light thereof will be suggested to persons skilled in the artand are included within the spirit and purview of this application andare considered within the scope of the appended claims. For example, therelative quantities of the ingredients may be varied to optimize thedesired effects, additional ingredients may be added, and/or similaringredients may be substituted for one or more of the ingredientsdescribed. Additional advantageous features and functionalitiesassociated with the systems, methods, and processes of the presentdisclosure will be apparent from the appended claims. Moreover, thoseskilled in the art will recognize, or be able to ascertain using no morethan routine experimentation, many equivalents to the specificembodiments of the disclosure described herein. Such equivalents areintended to be encompassed by the following claims.

What is claimed is:
 1. A compound having the chemical structure:PTM-L-CLM, or a pharmaceutically acceptable salt thereof, wherein: (a)the CLM is represented by the chemical structure:

wherein: W is selected from CH₂, O, CHR, C═O, SO₂, NH, N, optionallysubstituted cyclopropyl group, optionally substituted cyclobutyl group,and N-alkyl; W₃ is C or N; each X is independently selected from absent,O, S, and CH₂, Y is selected from CH₂, —C═CR′, NH, N-alkyl, N-aryl,N-heteroaryl, N-cycloalkyl, N-heterocyclyl, O, and S; Z is selected fromabsent, O, S, and CH₂; G and G′ are independently selected from H,unsubstituted or substituted linear or branched alkyl, OH, R′OCOOR,R′OCONRR″, CH₂-heterocyclyl optionally substituted with R′, and benzyloptionally substituted with R′; Q₁, Q₂, Q₃, and Q₄ represent C or Nsubstituted with a group independently selected from H, R, N, andN-oxide; A is independently selected from H, unsubstituted orsubstituted linear or branched alkyl, cycloalkyl, —Cl, and —F; m′ is 1,2, 3, 4, 5, 6, 7, 8, 9, or 10; R is a bond, H, —CONR′R″, —C(═O)R′, —OR′,—NR′R″, —SR′, —SO₂R′, —SO₂NR′R″, —CR′R″—, —CR′NR′R″—, (—CR′O)_(n′)R″,optionally substituted heterocyclyl, aryl, optionally substitutedalkyl-aryl, heteroaryl, unsubstituted or substituted linear or branchedalkyl, optionally substituted alkoxyl group, optionally substitutedheterocyclyl, —P(O)(OR′)R″, —P(O)R′R″, —OP(O)(OR′)R″, —OP(O)R′R″, —Cl,—F, —Br, —I, —CF₃, —CN, —NR′SO₂NR′R″, —NR′CONR′R″, —CONR′COR″,—NR′C(═N—CN)NR′R″, —C(═N—CN)NR′R″, —NR′C(═N—CN)R″, —NR′C(═C—NO₂)NR′R″,—SO₂NR′COR″, —NO₂, —CO₂R′, —C(C═N—OR′)R″, —CR′═CR′R″, —CCR′,—S(C═O)(C═N—R′)R″, —SF₅ and —OCF₃, wherein at least one R or W ismodified to be covalently joined to the PTM, or the CLM; R′ and R″ areindependently selected from a bond, H, optionally substituted alkyl,optionally substituted cycloalkyl, optionally substituted aryl,optionally substituted heteroaryl, optionally substituted heterocyclic,and optionally substituted heterocyclyl; n′ is 1, 2, 3, 4, 5, 6, 7, 8,9, or 10;

represents a single bond or a double bond; and

represents a bond that may be stereospecific or non-stereospecific; (b)the PTM has a chemical structure selected from:

wherein the

of the PTM indicates the site of attachment with a chemical linkinggroup; and (c) the L is a bond or is a chemical linking group thatcovalently couples the CLM to the PTM and is selected from:

wherein: N* is a nitrogen atom that is covalently linked to the CLM orthe PTM, or that is shared with the CLM or the PTM; each m, n, o, and pis independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, or 20; and the chemical linker group is optionallysubstituted with 1, 2, 3, or 4 substitutions independently selected fromhalogen and C₁₋₄ alkyl.
 2. The compound according to claim 1, whereinPTM is:

wherein

indicates a site of attachment of L or the CLM, and wherein each PTM iscoupled to at least one L or the CLM.
 3. The compound of claim 1,wherein the CLM is:


4. The compound of claim 1, wherein the CLM is:


5. The compound of to claim 1, wherein PTM is:

wherein

indicates a site of attachment of L or the CLM, and wherein each PTM iscoupled to at least one or the CLM.
 6. The compound claim 1, wherein:(a) the CLM is:

 wherein:

of the CLM indicates the point of attachment with L or the PTM; and N*is a nitrogen atom that is shared with the L or the PTM; (b) the PTM is:

 wherein

of the PTM indicates the point of attachment with L or the CLM; (c) L isselected from:

 wherein

or * indicates the site that is covalently linked to the CLM or the PTM,or that is shared with the CLM or the PTM; or (d) a combination thereof.7. The compound of claim 1, wherein: (a) the CLM is:

 wherein:

of the CLM indicates the point of attachment with L or the PTM; and N*is a nitrogen atom that is shared with L or the PTM; (b) the PTM is:

 wherein

of the PTM indicates the point of attachment with L or the CLM; and (c)L is selected from:

 wherein

or * indicates the site that is covalently linked to the CLM or the PTM,or that is shared with the CLM or the PTM.
 8. The compound of claim 1,wherein the compound is selected from:


9. A pharmaceutical composition comprising an amount of a compound ofclaim 1, and a pharmaceutically acceptable carrier.
 10. Thepharmaceutical composition of claim 9, wherein the composition furthercomprises an additional bioactive agent.
 11. The pharmaceuticalcomposition of claim 10, wherein the additional bioactive agent is ananti-neurodegenerative, an anti-inflammatory, a chemotherapy agent, oran immunomodulatory agent.
 12. The compound of claim 1, wherein thecompound is:

or a pharmaceutically acceptable salt thereof.